%PDF-1.6
%
1 0 obj
<>
endobj
2 0 obj
<>stream
Ekaterina K. Donchenko, Evgeniya V. Pechnikova, Maryia Yu. Mishyna, Tatiana I. Manukhova, Olga S. Sokolova, Nikolai A. Nikitin, Joseph G. Atabekov, Olga V. Karpova
Structure and properties of virions and virus-like particles derived from the coat protein of Alternanthera mosaic virus
www.plosone.org
www.plosone.org
endstream
endobj
3 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 12 0 R/Annots 13 0 R/Contents 14 0 R/TrimBox[0 0 612 792]>>
endobj
13 0 obj
[15 0 R 16 0 R 17 0 R 18 0 R 19 0 R 20 0 R 21 0 R 22 0 R 23 0 R 24 0 R 25 0 R 26 0 R 27 0 R 28 0 R]
endobj
15 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref011)>>
endobj
16 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref012)>>
endobj
17 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref013)>>
endobj
18 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref014)>>
endobj
19 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref015)>>
endobj
20 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref015)>>
endobj
21 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref016)>>
endobj
22 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref019)>>
endobj
23 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref020)>>
endobj
24 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref019)>>
endobj
25 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref021)>>
endobj
26 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref023)>>
endobj
27 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref010)>>
endobj
28 0 obj
<>/Border[0 0 0]/A 29 0 R>>
endobj
29 0 obj
<>
endobj
14 0 obj
[30 0 R 31 0 R 32 0 R 33 0 R 34 0 R 35 0 R 36 0 R 37 0 R 38 0 R 39 0 R 40 0 R 41 0 R 42 0 R 43 0 R]
endobj
30 0 obj
<>stream
q
0.83 0.64 0.02 0 k
287.0929 680.315 m
296.6173 680.315 l
h
f*
395.2063 680.315 m
404.7307 680.315 l
h
f*
513.411 680.315 m
522.8787 680.315 l
h
f*
311.2441 641.3102 m
320.7685 641.3102 l
h
f*
430.0157 628.3276 m
439.5402 628.3276 l
h
f*
439.2567 602.3055 m
448.7811 602.3055 l
h
f*
482.5134 576.3402 m
492.0378 576.3402 l
h
f*
497.1969 576.3402 m
506.7213 576.3402 l
h
f*
246.4441 550.3181 m
255.9118 550.3181 l
h
f*
274.3937 485.348 m
283.8614 485.348 l
h
f*
369.2976 472.3087 m
378.822 472.3087 l
h
f*
383.9811 472.3087 m
393.5055 472.3087 l
h
f*
246.4441 289.9276 m
255.9685 289.9276 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 211.9748 707.4141 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(In)Tj
1.1111 0 Td
(contrast)Tj
3.4639 0 Td
(to)Tj
1.0262 0 Td
(rich)Tj
1.8085 0 Td
(information)Tj
5.074 0 Td
(on)Tj
1.2755 0 Td
(the)Tj
1.4684 0 Td
(rigid)Tj
2.1203 0 Td
(rod)Tj
1.627 0 Td
(plant)Tj
2.2621 0 Td
(viruses,)Tj
3.2144 0 Td
(structural)Tj
4.0762 0 Td
(information)Tj
5.0797 0 Td
(for)Tj
-34.8035 -1.3039 Td
(flexible)Tj
3.084 0 Td
(plant)Tj
2.2677 0 Td
(viruses)Tj
2.982 0 Td
(has)Tj
1.5477 0 Td
(been)Tj
2.1204 0 Td
(lacking)Tj
3.101 0 Td
(for)Tj
1.3833 0 Td
(long)Tj
1.9899 0 Td
(time.)Tj
2.2507 0 Td
(Now,)Tj
2.3755 0 Td
(the)Tj
1.4683 0 Td
(high-resolution)Tj
6.4176 0 Td
(structures)Tj
4.1839 0 Td
(are)Tj
-35.172 -1.2982 Td
(available)Tj
3.6679 0 Td
(for)Tj
1.3833 0 Td
(PapMV)Tj
3.3166 0 Td
([)Tj
0.83 0.64 0.02 0 k
(11)Tj
0 g
(],)Tj
2.0806 0 Td
(bamboo)Tj
3.4979 0 Td
(mosaic)Tj
3.0331 0 Td
(virus)Tj
2.1996 0 Td
([)Tj
0.83 0.64 0.02 0 k
(12)Tj
0 g
(])Tj
1.8539 0 Td
(and)Tj
1.7291 0 Td
(pepino)Tj
2.999 0 Td
(mosaic)Tj
3.0331 0 Td
(virus)Tj
2.2053 0 Td
([)Tj
0.83 0.64 0.02 0 k
(13)Tj
0 g
(].)Tj
2.075 0 Td
(The)Tj
1.7801 0 Td
(low-)Tj
-34.8545 -1.2983 Td
(resolution)Tj
4.2746 0 Td
(structures)Tj
4.1839 0 Td
(for)Tj
1.3889 0 Td
(a)Tj
0.652 0 Td
(number)Tj
3.4186 0 Td
(of)Tj
1.0148 0 Td
(other)Tj
2.347 0 Td
(Potexviruses)Tj
5.2384 0 Td
(exist,)Tj
2.2621 0 Td
(but)Tj
1.5533 0 Td
(the)Tj
1.4684 0 Td
(3D)Tj
1.4343 0 Td
(structure)Tj
3.8211 0 Td
(of)Tj
1.0148 0 Td
(AltMV)Tj
-34.0722 -1.2983 Td
(virion)Tj
2.6361 0 Td
(has)Tj
1.5421 0 Td
(been)Tj
2.1203 0 Td
(lacking)Tj
3.1011 0 Td
(until)Tj
2.1089 0 Td
(now.)Tj
-10.3123 -1.3039 Td
(Similarly)Tj
3.804 0 Td
(to)Tj
1.0262 0 Td
(PapMV)Tj
3.3165 0 Td
(CP)Tj
1.4456 0 Td
([)Tj
0.83 0.64 0.02 0 k
(14)Tj
0 g
(],)Tj
2.075 0 Td
(AltMV)Tj
3.033 0 Td
(CP)Tj
1.4457 0 Td
(polymerizes)Tj
5.023 0 Td
(to)Tj
1.0261 0 Td
(form)Tj
2.2053 0 Td
(extended)Tj
3.8551 0 Td
(RNA-free)Tj
4.1216 0 Td
(virus-like)Tj
-33.5733 -1.2982 Td
(particles)Tj
3.5602 0 Td
(\(VLPs\))Tj
/F8 1 Tf
3.0558 0 Td
[()12()]TJ
1.0148 0 Td
[()-13()14()7()20()]TJ
/F7 1 Tf
2.0409 0 Td
(at)Tj
0.9524 0 Td
(pH)Tj
1.508 0 Td
(4.0)Tj
1.3947 0 Td
(and)Tj
1.7291 0 Td
(low)Tj
1.6611 0 Td
(ionic)Tj
2.2337 0 Td
(strength)Tj
3.5093 0 Td
([)Tj
0.83 0.64 0.02 0 k
(15)Tj
0 g
(].)Tj
2.0806 0 Td
(However,)Tj
4.0762 0 Td
(the)Tj
1.4683 0 Td
(PapMV)Tj
3.3165 0 Td
(CP)Tj
1.4457 0 Td
(does)Tj
-35.0473 -1.2983 Td
(not)Tj
1.5703 0 Td
(form)Tj
2.2054 0 Td
(VLPs)Tj
2.3754 0 Td
(at)Tj
0.9524 0 Td
(pH)Tj
1.5024 0 Td
(8.0)Tj
1.4003 0 Td
(in)Tj
1.0318 0 Td
(the)Tj
1.474 0 Td
(absence)Tj
3.3392 0 Td
(of)Tj
1.0148 0 Td
(RNA,)Tj
2.4888 0 Td
(while)Tj
2.3697 0 Td
(the)Tj
1.4684 0 Td
(AltMV)Tj
3.033 0 Td
(CP)Tj
1.4457 0 Td
(under)Tj
2.6192 0 Td
(these)Tj
2.2507 0 Td
(conditions)Tj
-32.5415 -1.3039 Td
(forms)Tj
2.5681 0 Td
(VLPs)Tj
2.3698 0 Td
(morphologically)Tj
6.7577 0 Td
(close)Tj
2.1827 0 Td
(to)Tj
1.0261 0 Td
(native)Tj
2.6476 0 Td
(AltMV)Tj
3.033 0 Td
(virions)Tj
2.999 0 Td
([)Tj
0.83 0.64 0.02 0 k
(15)Tj
0 g
(].)Tj
-22.3878 -1.2983 Td
(Recently,)Tj
3.8891 0 Td
(plant)Tj
2.262 0 Td
(viruses)Tj
2.9877 0 Td
(and)Tj
1.7291 0 Td
(VLPs,)Tj
2.5965 0 Td
(derived)Tj
3.2202 0 Td
(from)Tj
2.2053 0 Td
(them,)Tj
2.5115 0 Td
(have)Tj
2.0636 0 Td
(been)Tj
2.1203 0 Td
(used)Tj
2.0579 0 Td
(extensively)Tj
4.5865 0 Td
(in)Tj
1.0374 0 Td
(bio-)Tj
-34.4633 -1.2983 Td
(technology)Tj
4.6204 0 Td
(including)Tj
4.0422 0 Td
(the)Tj
1.4683 0 Td
(development)Tj
5.4085 0 Td
(of)Tj
1.0148 0 Td
(medical)Tj
3.3618 0 Td
(products)Tj
3.7587 0 Td
(\(vaccines\))Tj
4.235 0 Td
([)Tj
0.83 0.64 0.02 0 k
(16)Tj
0 g
()Tj
0.83 0.64 0.02 0 k
(19)Tj
0 g
(].)Tj
3.5433 0 Td
(Plant)Tj
2.3074 0 Td
(viruses)Tj
-33.7604 -1.3039 Td
(are)Tj
1.4456 0 Td
(absolutely)Tj
4.2293 0 Td
(safe)Tj
1.7234 0 Td
(for)Tj
1.389 0 Td
(humans,)Tj
3.6623 0 Td
(due)Tj
1.7008 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(low)Tj
1.6554 0 Td
(risk)Tj
1.7178 0 Td
(of)Tj
1.0148 0 Td
(cross-contamination)Tj
8.4925 0 Td
(with)Tj
2.0013 0 Td
(mammalian)Tj
-31.5267 -1.2982 Td
(pathogens)Tj
4.3029 0 Td
([)Tj
0.83 0.64 0.02 0 k
(20)Tj
0 g
(].)Tj
2.0749 0 Td
(Usage)Tj
2.6419 0 Td
(of)Tj
1.0148 0 Td
(plant)Tj
2.2621 0 Td
(viruses)Tj
2.9877 0 Td
(as)Tj
1.0147 0 Td
(a)Tj
0.652 0 Td
(basis)Tj
2.1487 0 Td
(for)Tj
1.3889 0 Td
(medical)Tj
3.3619 0 Td
(nanotechnologies)Tj
7.2566 0 Td
(reduces)Tj
3.2769 0 Td
(many)Tj
-34.384 -1.2983 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(risks)Tj
2.0749 0 Td
(associated)Tj
4.2576 0 Td
(with)Tj
1.9956 0 Td
(other)Tj
2.3527 0 Td
(biological)Tj
4.0876 0 Td
(materials,)Tj
4.0989 0 Td
(being)Tj
2.4321 0 Td
(emplemented)Tj
5.6919 0 Td
(in)Tj
1.0375 0 Td
(vaccine,)Tj
3.4185 0 Td
(as)Tj
1.0148 0 Td
(well)Tj
1.8199 0 Td
(as)Tj
-36.7651 -1.3039 Td
(the)Tj
1.4683 0 Td
(use)Tj
1.5307 0 Td
(of)Tj
1.0148 0 Td
(noninfective)Tj
5.2214 0 Td
(VLPs)Tj
2.3697 0 Td
(results)Tj
2.812 0 Td
(in)Tj
1.0318 0 Td
(low)Tj
1.6611 0 Td
(risk)Tj
1.7121 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.474 0 Td
(environment)Tj
5.4255 0 Td
(and)Tj
1.7348 0 Td
(human)Tj
3.0784 0 Td
(beings.)Tj
3.016 0 Td
(These)Tj
-34.5767 -1.2983 Td
(lower)Tj
2.4491 0 Td
(risks)Tj
2.0806 0 Td
(allow)Tj
2.3357 0 Td
(for)Tj
1.3833 0 Td
(easier)Tj
2.5001 0 Td
(handling,)Tj
4.0139 0 Td
(transporting)Tj
5.21 0 Td
(and)Tj
1.7291 0 Td
(processing)Tj
4.4618 0 Td
(of)Tj
1.0147 0 Td
(viral)Tj
1.9956 0 Td
(nanoparticles,)Tj
5.828 0 Td
(mak-)Tj
-35.0019 -1.2982 Td
(ing)Tj
1.4966 0 Td
(plant)Tj
2.2677 0 Td
(virus-based)Tj
4.8019 0 Td
(particles)Tj
3.5659 0 Td
(particularly)Tj
4.8076 0 Td
(attractive)Tj
3.9231 0 Td
(platforms)Tj
4.0649 0 Td
(for)Tj
1.3833 0 Td
(a)Tj
0.6576 0 Td
(range)Tj
2.4604 0 Td
(of)Tj
1.0148 0 Td
(nanobiotechnolo-)Tj
-30.4438 -1.2983 Td
(gical)Tj
2.0522 0 Td
(applications)Tj
5.0456 0 Td
([)Tj
0.83 0.64 0.02 0 k
(19)Tj
0 g
(].)Tj
2.075 0 Td
(Virions)Tj
3.2371 0 Td
(and)Tj
1.7292 0 Td
(VLPs)Tj
2.3754 0 Td
(of)Tj
1.0148 0 Td
(PapMV)Tj
3.3165 0 Td
(\(closely)Tj
3.2201 0 Td
(related)Tj
2.948 0 Td
(to)Tj
1.0205 0 Td
(AltMV\))Tj
3.3789 0 Td
(were)Tj
2.1203 0 Td
(shown)Tj
2.8516 0 Td
(to)Tj
-36.3852 -1.3039 Td
(have)Tj
2.0579 0 Td
(high)Tj
2.0069 0 Td
(immunostimulating)Tj
8.2431 0 Td
(properties)Tj
4.2803 0 Td
([)Tj
0.83 0.64 0.02 0 k
(21)Tj
0 g
()Tj
0.83 0.64 0.02 0 k
(23)Tj
0 g
(].)Tj
3.5433 0 Td
(Thereby,)Tj
3.7643 0 Td
(the)Tj
1.4684 0 Td
(study)Tj
2.3924 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(structure)Tj
3.8211 0 Td
(and)Tj
1.7348 0 Td
(sta-)Tj
-35.7956 -1.2983 Td
(bility)Tj
2.2563 0 Td
(of)Tj
1.0148 0 Td
(AltMV)Tj
3.0387 0 Td
(virions)Tj
2.9991 0 Td
(and)Tj
1.7291 0 Td
(VLPs)Tj
2.3697 0 Td
(can)Tj
1.6271 0 Td
(be)Tj
1.1509 0 Td
(a)Tj
0.6519 0 Td
(promising)Tj
4.3483 0 Td
(and)Tj
1.7292 0 Td
(tremendously)Tj
5.7259 0 Td
(important)Tj
4.286 0 Td
(direction)Tj
3.8551 0 Td
(of)Tj
-36.7821 -1.2983 Td
(research.)Tj
1.1962 -1.3039 Td
(This)Tj
1.9899 0 Td
(work)Tj
2.279 0 Td
(is)Tj
0.8447 0 Td
(devoted)Tj
3.3846 0 Td
(to)Tj
1.0261 0 Td
(investigation)Tj
5.3574 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.474 0 Td
(possibility)Tj
4.2747 0 Td
(of)Tj
1.0148 0 Td
(obtaining)Tj
4.0592 0 Td
(AltMV)Tj
3.0387 0 Td
(VLPs)Tj
2.3697 0 Td
(under)Tj
2.6192 0 Td
(dif-)Tj
-35.943 -1.2982 Td
(ferent)Tj
2.5738 0 Td
(conditions)Tj
4.473 0 Td
(and)Tj
1.7291 0 Td
(to)Tj
1.0262 0 Td
(a)Tj
0.6576 0 Td
(detailed)Tj
3.3676 0 Td
(characterization)Tj
6.6613 0 Td
(of)Tj
1.0148 0 Td
(AltMV)Tj
3.0331 0 Td
(virions)Tj
2.999 0 Td
(and)Tj
1.7291 0 Td
(VLPs)Tj
2.3755 0 Td
(properties)Tj
4.2746 0 Td
(in)Tj
-35.9147 -1.2983 Td
(terms)Tj
2.4887 0 Td
(of)Tj
1.0148 0 Td
(their)Tj
2.109 0 Td
(biotechnological)Tj
6.8144 0 Td
(potential.)Tj
3.9855 0 Td
(In)Tj
1.1055 0 Td
(addition,)Tj
3.8211 0 Td
(the)Tj
1.4684 0 Td
(method)Tj
3.3221 0 Td
(of)Tj
1.0148 0 Td
(AltMV)Tj
3.0331 0 Td
(isolation)Tj
3.6567 0 Td
(and)Tj
1.7291 0 Td
(puri-)Tj
-35.5632 -1.3039 Td
(fication)Tj
3.2541 0 Td
(was)Tj
1.6951 0 Td
(modified,)Tj
4.0762 0 Td
(markedly)Tj
3.9911 0 Td
(increasing)Tj
4.3314 0 Td
(the)Tj
1.4683 0 Td
(yield)Tj
2.143 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(purified)Tj
3.4072 0 Td
(virus.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 364.9889 cm
BT
/F2 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Materials)Tj
4.5071 0 Td
(and)Tj
1.989 0 Td
(methods)Tj
/F0 1 Tf
-6.4961 -1.4976 Td
(Virus)Tj
2.4708 0 Td
(purification)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 330.0094 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(The)Tj
1.7801 0 Td
(initial)Tj
2.5512 0 Td
(infected)Tj
3.4185 0 Td
(material)Tj
3.515 0 Td
(was)Tj
1.7007 0 Td
(received)Tj
3.532 0 Td
(from)Tj
2.2053 0 Td
(the)Tj
1.4684 0 Td
(German)Tj
3.5319 0 Td
(Collection)Tj
4.354 0 Td
(of)Tj
1.0148 0 Td
(Microorganisms)Tj
6.8144 0 Td
(and)Tj
-35.8863 -1.2982 Td
(Cell)Tj
1.7971 0 Td
(Cultures,)Tj
3.8664 0 Td
(Braunschweig)Tj
5.8337 0 Td
(as)Tj
1.0148 0 Td
(a)Tj
0.652 0 Td
(strain)Tj
2.5001 0 Td
(of)Tj
1.0148 0 Td
(PapMV.)Tj
3.5433 0 Td
(We)Tj
1.6101 0 Td
(propagated)Tj
4.7168 0 Td
(this)Tj
1.6724 0 Td
(virus)Tj
2.1997 0 Td
(and)Tj
1.7348 0 Td
(defined)Tj
3.2258 0 Td
(it)Tj
0.7823 0 Td
(as)Tj
-36.1641 -1.3039 Td
(a)Tj
0.6519 0 Td
(new)Tj
1.8765 0 Td
(strain)Tj
2.5002 0 Td
(of)Tj
1.0148 0 Td
(AltMVAltMV-MU)Tj
8.3508 0 Td
(\(Moscow)Tj
3.9345 0 Td
(University\))Tj
4.7395 0 Td
(\(accession)Tj
4.3086 0 Td
(number)Tj
3.4185 0 Td
(FJ822136)Tj
3.9345 0 Td
(in)Tj
1.0318 0 Td
(the)Tj
-35.7616 -1.2983 Td
(GenBank\))Tj
4.3029 0 Td
([)Tj
0.83 0.64 0.02 0 k
(10)Tj
0 g
(].)Tj
-3.1067 -1.2983 Td
(The)Tj
1.7858 0 Td
(virus)Tj
2.1996 0 Td
(was)Tj
1.7008 0 Td
(maintained)Tj
4.7905 0 Td
(in)Tj
1.0319 0 Td
(either)Tj
/F8 1 Tf
2.5341 0 Td
[()35()4()7()12()7()4()16()6()]TJ
4.0819 0 Td
[()4()2()22()10()6()16()18()4()16()6()]TJ
/F7 1 Tf
5.4538 0 Td
(or)Tj
/F8 1 Tf
1.0942 0 Td
[()23()11()-19()7( )4()4()6()7()]TJ
4.0025 0 Td
[()-8()20()16()6()12()12()-36()4()11()20()]TJ
/F7 1 Tf
(.)Tj
4.8132 0 Td
(Plants)Tj
-34.6845 -1.3039 Td
(were)Tj
2.1203 0 Td
(provided)Tj
3.8154 0 Td
(by)Tj
1.1848 0 Td
(the)Tj
1.4684 0 Td
(greenhouse)Tj
4.7962 0 Td
(of)Tj
1.0148 0 Td
(Faculty)Tj
3.1351 0 Td
(of)Tj
1.0148 0 Td
(Biology,)Tj
3.4695 0 Td
(Lomonosov)Tj
4.972 0 Td
(Moscow)Tj
3.5943 0 Td
(State)Tj
2.1486 0 Td
(University.)Tj
-32.7342 -1.2982 Td
(Plants)Tj
2.6645 0 Td
(were)Tj
2.126 0 Td
(inoculated)Tj
4.422 0 Td
(using)Tj
2.3867 0 Td
(sap)Tj
1.5364 0 Td
(extracts)Tj
3.2939 0 Td
(\(c.)Tj
1.2075 0 Td
(1:51:10,)Tj
3.7927 0 Td
(w:v)Tj
1.5818 0 Td
(of)Tj
1.0148 0 Td
(0.01)Tj
1.8765 0 Td
(M)Tj
1.1112 0 Td
(Tris-HCl,)Tj
4.0818 0 Td
(pH)Tj
1.508 0 Td
(7.5,)Tj
1.6214 0 Td
(with)Tj
-34.2252 -1.2983 Td
(Celite)Tj
2.5398 0 Td
(added)Tj
2.6589 0 Td
(as)Tj
1.0148 0 Td
(an)Tj
1.2075 0 Td
(abrasive\))Tj
3.8097 0 Td
(from)Tj
2.2054 0 Td
(plant)Tj
2.2677 0 Td
(tissue)Tj
2.4548 0 Td
(of)Tj
1.0148 0 Td
(infected)Tj
/F8 1 Tf
3.4242 0 Td
[()]TJ
/F7 1 Tf
(.)Tj
/F8 1 Tf
0.9921 0 Td
[()-8()20()16()6()12()12()-36()4()11()20()]TJ
/F7 1 Tf
(.)Tj
4.8132 0 Td
(30)Tj
1.1679 0 Td
(days)Tj
2.0012 0 Td
(after)Tj
2.0353 0 Td
(infection,)Tj
-33.6073 -1.3039 Td
(the)Tj
1.4683 0 Td
(plant)Tj
2.262 0 Td
(biomass)Tj
3.4809 0 Td
(was)Tj
1.6952 0 Td
(frozen)Tj
2.7722 0 Td
(at)Tj
0.9525 0 Td
(-20C.)Tj
2.8006 0 Td
(For)Tj
1.627 0 Td
(virus)Tj
2.2054 0 Td
(purification,)Tj
5.159 0 Td
(plants)Tj
2.6249 0 Td
(were)Tj
2.1259 0 Td
(homogenized)Tj
5.6466 0 Td
(in)Tj
1.0375 0 Td
(0.3)Tj
-35.858 -1.2983 Td
(M)Tj
1.1111 0 Td
(glycine-KOH,)Tj
5.811 0 Td
(1%)Tj
1.457 0 Td
(Na2SO3,)Tj
3.7871 0 Td
(pH)Tj
1.5023 0 Td
(7.5)Tj
1.4003 0 Td
(\(3)Tj
1.0318 0 Td
(ml)Tj
1.287 0 Td
(buffer)Tj
2.6248 0 Td
(per)Tj
1.5364 0 Td
(1)Tj
0.6916 0 Td
(g)Tj
0.686 0 Td
(of)Tj
1.0148 0 Td
(leaves\))Tj
2.9027 0 Td
(in)Tj
1.0374 0 Td
(a)Tj
0.652 0 Td
(blender.)Tj
3.4923 0 Td
(All)Tj
1.4003 0 Td
(steps)Tj
2.177 0 Td
(of)Tj
-35.6029 -1.2983 Td
(virus)Tj
2.1996 0 Td
(preparations)Tj
5.2724 0 Td
(were)Tj
2.1203 0 Td
(carried)Tj
3.0388 0 Td
(out)Tj
1.5533 0 Td
(at)Tj
0.9525 0 Td
(4C)Tj
1.7518 0 Td
(and)Tj
1.7291 0 Td
(with)Tj
1.9956 0 Td
(chilled)Tj
2.8856 0 Td
(buffers.)Tj
3.2088 0 Td
(The)Tj
1.7858 0 Td
(plant)Tj
2.2621 0 Td
(sap)Tj
1.5307 0 Td
(was)Tj
1.7007 0 Td
(then)Tj
2.0183 0 Td
(fil-)Tj
-36.0054 -1.3039 Td
(tered)Tj
2.262 0 Td
(through)Tj
3.4469 0 Td
(a)Tj
0.652 0 Td
(cheesecloth,)Tj
5.0343 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4683 0 Td
(filtrate)Tj
2.846 0 Td
(centrifuged)Tj
4.7848 0 Td
(at)Tj
0.9525 0 Td
(12,000)Tj
2.8233 0 Td
(rpm)Tj
1.9275 0 Td
(for)Tj
1.389 0 Td
(30)Tj
1.1678 0 Td
(min)Tj
1.8596 0 Td
(\(Beckman)Tj
-32.3431 -1.2982 Td
(J2-21)Tj
2.3243 0 Td
(Centrifuge,)Tj
4.7225 0 Td
(JA-14)Tj
2.5342 0 Td
(rotor\).)Tj
2.8403 0 Td
(The)Tj
1.7858 0 Td
(supernatant)Tj
4.9833 0 Td
(was)Tj
1.6951 0 Td
(collected)Tj
3.7417 0 Td
(and)Tj
1.7291 0 Td
(clarified)Tj
3.4696 0 Td
(with)Tj
1.9956 0 Td
(Triton)Tj
2.8233 0 Td
(X-100)Tj
-34.6448 -1.2983 Td
(added)Tj
2.6588 0 Td
(to)Tj
1.0262 0 Td
(1%)Tj
1.4513 0 Td
(\(v/v\))Tj
2.143 0 Td
(and)Tj
1.7291 0 Td
(incubated)Tj
4.1782 0 Td
(for)Tj
1.3833 0 Td
(20)Tj
1.1736 0 Td
(min.)Tj
2.0806 0 Td
(Polyethylene)Tj
5.3291 0 Td
(glycol)Tj
2.5625 0 Td
(\(PEG;)Tj
2.6305 0 Td
(MW)Tj
2.0806 0 Td
(6000\))Tj
2.4605 0 Td
(and)Tj
1.7291 0 Td
(NaCl)Tj
-34.6164 -1.2983 Td
(were)Tj
2.1203 0 Td
(added)Tj
2.6588 0 Td
(to)Tj
1.0262 0 Td
(the)Tj
1.474 0 Td
(solution)Tj
3.4809 0 Td
(to)Tj
1.0261 0 Td
(5%)Tj
1.4514 0 Td
(and)Tj
1.7291 0 Td
(to)Tj
1.0261 0 Td
(2%)Tj
1.4513 0 Td
(\(w/v\),)Tj
2.5966 0 Td
(respectively.)Tj
5.108 0 Td
(After)Tj
2.2847 0 Td
(dissolving)Tj
4.2292 0 Td
(the)Tj
1.4684 0 Td
(compo-)Tj
-33.1311 -1.3039 Td
(nents,)Tj
2.6248 0 Td
(the)Tj
1.4684 0 Td
(solution)Tj
3.4809 0 Td
(was)Tj
1.7008 0 Td
(kept)Tj
1.9558 0 Td
(overnight)Tj
4.0819 0 Td
(at)Tj
0.9524 0 Td
(+4C.)Tj
2.5569 0 Td
(The)Tj
1.7858 0 Td
(PEG)Tj
2.0636 0 Td
(precipitate)Tj
4.4617 0 Td
(was)Tj
1.6951 0 Td
(collected)Tj
3.7417 0 Td
(by)Tj
1.1792 0 Td
(centrifu-)Tj
-33.749 -1.2982 Td
(gation)Tj
2.7382 0 Td
(at)Tj
0.9524 0 Td
(10,000)Tj
2.8233 0 Td
(rpm)Tj
1.9276 0 Td
(for)Tj
1.3889 0 Td
(20)Tj
1.1679 0 Td
(min)Tj
1.8595 0 Td
(\(Beckman)Tj
4.286 0 Td
(J2-21)Tj
2.3244 0 Td
(Centrifuge,)Tj
4.7225 0 Td
(JA-14)Tj
2.5341 0 Td
(rotor\).)Tj
2.8403 0 Td
(The)Tj
1.7858 0 Td
(sediment)Tj
3.8891 0 Td
(was)Tj
-35.24 -1.2983 Td
(collected)Tj
3.736 0 Td
(and)Tj
1.7348 0 Td
(retained)Tj
3.5093 0 Td
(at)Tj
0.9524 0 Td
(+4C.)Tj
2.5568 0 Td
(The)Tj
1.7858 0 Td
(PEG)Tj
2.0636 0 Td
(\(Mr)Tj
1.8255 0 Td
(6000\))Tj
2.4605 0 Td
(was)Tj
1.7008 0 Td
(added)Tj
2.6589 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4683 0 Td
(supernatant)Tj
4.9833 0 Td
(to)Tj
1.0261 0 Td
(a)Tj
0.652 0 Td
(final)Tj
-34.1402 -1.3039 Td
(concentration)Tj
5.8166 0 Td
(of)Tj
1.0148 0 Td
(8%)Tj
1.4513 0 Td
(\(taking)Tj
3.0728 0 Td
(into)Tj
1.8425 0 Td
(account)Tj
3.3789 0 Td
(the)Tj
1.474 0 Td
(one)Tj
1.6951 0 Td
(added)Tj
2.6645 0 Td
(earlier\).)Tj
3.3222 0 Td
(After)Tj
2.2847 0 Td
(dissolving)Tj
4.2236 0 Td
(the)Tj
1.4683 0 Td
(com-)Tj
-33.7093 -1.2983 Td
(ponents,)Tj
3.651 0 Td
(the)Tj
1.4683 0 Td
(solution)Tj
3.4809 0 Td
(was)Tj
1.7008 0 Td
(kept)Tj
1.9559 0 Td
(overnight)Tj
4.0818 0 Td
(at)Tj
0.9525 0 Td
(+4C.)Tj
2.5568 0 Td
(The)Tj
1.7802 0 Td
(PEG)Tj
2.0636 0 Td
(precipitate)Tj
4.4617 0 Td
(was)Tj
1.7008 0 Td
(collected)Tj
3.7417 0 Td
(by)Tj
ET
Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 107.1496 23.6976 re
W* n
q
107.0929 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
Q
Q
q
0 0 612 792 re
W* n
1 j
1 J
0 w
7.9999 0 0 7.9999 390.2173 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(Structure)Tj
4.2165 0 Td
(and)Tj
1.8567 0 Td
(properties)Tj
4.578 0 Td
(of)Tj
1.0418 0 Td
(Alternanthe)Tj
5.0244 0 Td
(ra)Tj
1.0914 0 Td
(mosaic)Tj
3.3378 0 Td
(virus)Tj
ET
Q
q
1 j
1 J
0 w
36 48.0002 m
576 48.0002 l
576 47.5002 l
36 47.5002 l
f*
0.83 0.64 0.02 0 k
81.5811 34.9228 m
237.8835 34.9228 l
h
f*
0 g
7.9999 0 0 7.9999 36 36 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(PLOS)Tj
2.8559 0 Td
(ONE)Tj
2.3669 0 Td
(|)Tj
0.83 0.64 0.02 0 k
0.4748 0 Td
(https://doi.or)Tj
5.4142 0 Td
(g/10.137)Tj
3.8198 0 Td
(1/journal.po)Tj
5.1236 0 Td
(ne.01838)Tj
4.089 0 Td
(24)Tj
0 g
2.0906 0 Td
(August)Tj
3.2882 0 Td
(24,)Tj
1.5874 0 Td
(2017)Tj
34.0302 0 Td
(2)Tj
0.7654 0 Td
(/)Tj
0.496 0 Td
(11)Tj
/F10 1 Tf
-0.025 Tc
-66.402 84.0904 Td
(is)Tj
0.8362 0 Td
(employe)Tj
3.1535 0 Td
(d)Tj
0.6945 0 Td
(by)Tj
1.1197 0 Td
(a)Tj
0.6378 0 Td
(commercial)Tj
4.578 0 Td
(company)Tj
3.6638 0 Td
(`Thermo)Tj
-14.6835 -1.3748 Td
(Fisher)Tj
2.459 0 Td
(Scientific')Tj
3.5434 0 Td
(.)Tj
0.4464 0 Td
(The)Tj
1.5874 0 Td
(presented)Tj
3.8906 0 Td
(cryo-EM)Tj
3.3733 0 Td
(results)Tj
-15.3001 -1.3749 Td
(were)Tj
2.0126 0 Td
(obtained)Tj
3.4016 0 Td
(in)Tj
0.8858 0 Td
(V.A.)Tj
1.7221 0 Td
(Shoubniko)Tj
3.9756 0 Td
(v)Tj
0.6449 0 Td
(Institute)Tj
3.2244 0 Td
(of)Tj
-15.867 -1.3748 Td
(Crystallogr)Tj
3.9897 0 Td
(aphy)Tj
2.0127 0 Td
(RAS)Tj
1.8567 0 Td
(and)Tj
1.5874 0 Td
(National)Tj
3.2386 0 Td
(Research)Tj
3.6992 0 Td
(Centre)Tj
-16.3843 -1.3748 Td
(`Kurchatov)Tj
4.1882 0 Td
(Institute',)Tj
3.6354 0 Td
(where)Tj
2.4875 0 Td
(Pechnikova)Tj
4.4646 0 Td
(E.V.)Tj
1.6653 0 Td
(was)Tj
-16.441 -1.3748 Td
(employed)Tj
3.848 0 Td
(at)Tj
0.9 0 Td
(that)Tj
1.6229 0 Td
(time.)Tj
2.0693 0 Td
(`Thermo)Tj
3.3165 0 Td
(Fisher)Tj
2.4591 0 Td
(Scientific')Tj
-14.2158 -1.3748 Td
(provided)Tj
3.4582 0 Td
(support)Tj
3.0969 0 Td
(in)Tj
0.8859 0 Td
(the)Tj
1.3677 0 Td
(form)Tj
2.0126 0 Td
(of)Tj
0.9496 0 Td
(salary)Tj
2.4024 0 Td
(for)Tj
1.2614 0 Td
(author)Tj
-15.4347 -1.3748 Td
([Pechnikova)Tj
4.7764 0 Td
(E.V.],)Tj
2.1968 0 Td
(but)Tj
1.4174 0 Td
(did)Tj
1.3606 0 Td
(not)Tj
1.4244 0 Td
(have)Tj
1.9559 0 Td
(any)Tj
1.5379 0 Td
(additional)Tj
-14.6694 -1.3748 Td
(role)Tj
1.6157 0 Td
(in)Tj
0.8858 0 Td
(the)Tj
1.3678 0 Td
(study)Tj
2.2677 0 Td
(design,)Tj
2.8985 0 Td
(data)Tj
1.7929 0 Td
(collection)Tj
3.7417 0 Td
(and)Tj
-14.5701 -1.3748 Td
(analysis,)Tj
3.4086 0 Td
(decision)Tj
3.2953 0 Td
(to)Tj
0.9497 0 Td
(publish,)Tj
3.1393 0 Td
(or)Tj
1.0064 0 Td
(prepara)Tj
2.8205 0 Td
(tion)Tj
1.6157 0 Td
(of)Tj
0.9496 0 Td
(the)Tj
-17.1851 -1.3749 Td
(manuscript.)Tj
4.6417 0 Td
(The)Tj
1.5874 0 Td
(specific)Tj
3.019 0 Td
(role)Tj
1.6157 0 Td
(of)Tj
0.9496 0 Td
(this)Tj
1.5591 0 Td
(author)Tj
2.6292 0 Td
(is)Tj
-16.0017 -1.3748 Td
(articulated)Tj
4.0535 0 Td
(in)Tj
0.8929 0 Td
(the)Tj
1.3678 0 Td
(`author)Tj
2.8134 0 Td
(contributions')Tj
5.3008 0 Td
(section.)Tj
/F9 1 Tf
-14.4284 -2.126 Td
(Competing)Tj
4.3795 0 Td
(interests)Tj
3.3307 0 Td
(:)Tj
/F10 1 Tf
0.4819 0 Td
(Curren)Tj
2.5016 0 Td
(tly)Tj
1.0914 0 Td
(one)Tj
1.5874 0 Td
(of)Tj
0.9496 0 Td
(the)Tj
1.3748 0 Td
(authors)Tj
-15.6969 -1.3748 Td
(\(Pechnikova)Tj
4.7764 0 Td
(E.V.\))Tj
1.9701 0 Td
(is)Tj
0.8362 0 Td
(employed)Tj
3.848 0 Td
(by)Tj
1.1197 0 Td
(a)Tj
0.6378 0 Td
(commercial)Tj
-13.1882 -1.3748 Td
(company)Tj
3.6567 0 Td
(`Thermo)Tj
3.3165 0 Td
(Fisher)Tj
2.4591 0 Td
(Scientific')Tj
3.5433 0 Td
(.)Tj
0.4465 0 Td
(The)Tj
1.5874 0 Td
(presented)Tj
-15.0095 -1.3748 Td
(cryo-EM)Tj
3.3803 0 Td
(results)Tj
2.7142 0 Td
(were)Tj
2.0055 0 Td
(obtained)Tj
3.4087 0 Td
(in)Tj
0.8858 0 Td
(V.A.)Tj
1.715 0 Td
(Shoubnikov)Tj
-14.1095 -1.3748 Td
(Institute)Tj
3.2244 0 Td
(of)Tj
0.9496 0 Td
(Crystallogr)Tj
3.9898 0 Td
(aphy)Tj
2.0126 0 Td
(RAS)Tj
1.8567 0 Td
(and)Tj
1.5874 0 Td
(National)Tj
-13.6205 -1.3748 Td
(Research)Tj
3.6992 0 Td
(Centre)Tj
2.6221 0 Td
(`Kurchatov)Tj
4.1811 0 Td
(Institute',)Tj
3.6425 0 Td
(where)Tj
-14.1449 -1.3749 Td
(Pechnikova)Tj
4.4646 0 Td
(E.V.)Tj
1.6653 0 Td
(was)Tj
1.7079 0 Td
(employe)Tj
3.1536 0 Td
(d)Tj
0.6945 0 Td
(at)Tj
0.9 0 Td
(that)Tj
1.6228 0 Td
(time.)Tj
2.0693 0 Td
(This)Tj
-16.278 -1.3748 Td
(current)Tj
2.8771 0 Td
(affiliation)Tj
3.5505 0 Td
(of)Tj
0.9496 0 Td
(Pechnikova)Tj
4.4646 0 Td
(E.V.)Tj
1.6653 0 Td
(does)Tj
2.0127 0 Td
(not)Tj
1.4244 0 Td
(alter)Tj
-16.9442 -1.3748 Td
(our)Tj
1.474 0 Td
(adherence)Tj
4.0536 0 Td
(to)Tj
0.9496 0 Td
(PLOS)Tj
2.3315 0 Td
(ONE)Tj
1.8709 0 Td
(policies)Tj
3.0118 0 Td
(on)Tj
1.1622 0 Td
(sharing)Tj
-14.8536 -1.3748 Td
(data)Tj
1.7929 0 Td
(and)Tj
1.5874 0 Td
(materials.)Tj
/F9 1 Tf
-3.3803 -2.126 Td
(Abbreviati)Tj
3.9118 0 Td
(ons:)Tj
/F10 1 Tf
1.8425 0 Td
(AltMV,)Tj
2.7 0 Td
(Alternanther)Tj
4.5284 0 Td
(a)Tj
0.6449 0 Td
(mosaic)Tj
2.8985 0 Td
(virus;)Tj
-16.5261 -1.3748 Td
(CP,)Tj
1.4952 0 Td
(coat)Tj
1.793 0 Td
(protein;)Tj
3.0401 0 Td
(EM,)Tj
1.6725 0 Td
(electron)Tj
3.189 0 Td
(microscop)Tj
3.9331 0 Td
(y;)Tj
0.8646 0 Td
(FSC,)Tj
-15.9875 -1.3748 Td
(Fourier)Tj
2.8204 0 Td
(shell)Tj
1.9205 0 Td
(correlation;)Tj
4.3867 0 Td
(PapMV)Tj
2.6929 0 Td
(,)Tj
0.4465 0 Td
(papaya)Tj
2.8559 0 Td
(mosaic)Tj
-15.1229 -1.3748 Td
(virus;)Tj
2.2606 0 Td
(TEM,)Tj
2.1473 0 Td
(transmi)Tj
2.8205 0 Td
(ssion)Tj
2.2039 0 Td
(electron)Tj
3.189 0 Td
(microscopy)Tj
4.3583 0 Td
(;)Tj
-16.9796 -1.3749 Td
(VLP,)Tj
1.97 0 Td
(virus-lik)Tj
2.9268 0 Td
(e)Tj
ET
endstream
endobj
31 0 obj
<>stream
endstream
endobj
32 0 obj
<>stream
endstream
endobj
33 0 obj
<>stream
endstream
endobj
34 0 obj
<>stream
endstream
endobj
35 0 obj
<>stream
endstream
endobj
36 0 obj
<>stream
endstream
endobj
37 0 obj
<>stream
endstream
endobj
38 0 obj
<>stream
endstream
endobj
39 0 obj
<>stream
endstream
endobj
40 0 obj
<>stream
endstream
endobj
41 0 obj
<>stream
endstream
endobj
42 0 obj
<>stream
endstream
endobj
43 0 obj
<>stream
BT
5.5417 46.8428 Td
(particle.)Tj
ET
Q
endstream
endobj
44 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 12 0 R/Annots 46 0 R/Contents 47 0 R/TrimBox[0 0 612 792]>>
endobj
46 0 obj
[48 0 R 49 0 R 50 0 R 51 0 R 52 0 R 53 0 R 54 0 R]
endobj
48 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref001)>>
endobj
49 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref010)>>
endobj
50 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref010)>>
endobj
51 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref024)>>
endobj
52 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref025)>>
endobj
53 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref026)>>
endobj
54 0 obj
<>/Border[0 0 0]/A 55 0 R>>
endobj
55 0 obj
<>
endobj
47 0 obj
[56 0 R 57 0 R 58 0 R 59 0 R 60 0 R 61 0 R 62 0 R 63 0 R 64 0 R 65 0 R 66 0 R]
endobj
56 0 obj
<>stream
q
0.83 0.64 0.02 0 k
513.411 537.3354 m
518.1732 537.3354 l
h
f*
520.4409 537.3354 m
529.9654 537.3354 l
h
f*
497.9906 480.0189 m
507.515 480.0189 l
h
f*
509.726 480.0189 m
519.2504 480.0189 l
h
f*
428.485 365.4992 m
438.0094 365.4992 l
h
f*
449.1213 94.9039 m
458.6457 94.9039 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 200.0125 707.4141 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(centrifugation)Tj
5.8846 0 Td
(at)Tj
0.9525 0 Td
(10,000)Tj
2.8233 0 Td
(rpm)Tj
1.9332 0 Td
(for)Tj
1.3833 0 Td
(20)Tj
1.1735 0 Td
(min)Tj
1.8539 0 Td
(\(Beckman)Tj
4.2859 0 Td
(J2-21)Tj
2.3244 0 Td
(Centrifuge,)Tj
4.7225 0 Td
(JA-14)Tj
2.5342 0 Td
(rotor\))Tj
2.6192 0 Td
(and)Tj
1.7291 0 Td
(com-)Tj
-34.2196 -1.3039 Td
(bined)Tj
2.4944 0 Td
(with)Tj
1.9956 0 Td
(the)Tj
1.4683 0 Td
(virus)Tj
2.2054 0 Td
(precipitate)Tj
4.4617 0 Td
(obtained)Tj
3.7304 0 Td
(on)Tj
1.2755 0 Td
(the)Tj
1.4684 0 Td
(previous)Tj
3.651 0 Td
(day.)Tj
-21.5545 -1.2982 Td
(Extraction)Tj
4.405 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(obtained)Tj
3.736 0 Td
(precipitate)Tj
4.4618 0 Td
(was)Tj
1.6951 0 Td
(carried)Tj
3.0387 0 Td
(out)Tj
1.5534 0 Td
(two)Tj
1.7121 0 Td
(to)Tj
1.0261 0 Td
(three)Tj
2.262 0 Td
(times)Tj
2.3925 0 Td
(for)Tj
1.3833 0 Td
(26)Tj
1.6894 0 Td
(hours)Tj
2.5115 0 Td
(by)Tj
-35.5462 -1.2983 Td
(gentle)Tj
2.6248 0 Td
(shaking)Tj
3.3165 0 Td
(\(until)Tj
2.4491 0 Td
(the)Tj
1.4741 0 Td
(precipitate)Tj
4.456 0 Td
(was)Tj
1.7008 0 Td
(completely)Tj
4.5807 0 Td
(dissolved\))Tj
4.235 0 Td
(with)Tj
1.9955 0 Td
(a)Tj
0.6577 0 Td
(buffer)Tj
2.6248 0 Td
(containing)Tj
4.5298 0 Td
(0.05)Tj
1.8708 0 Td
(M)Tj
-36.5156 -1.2983 Td
(Tris-HC1,)Tj
4.3143 0 Td
(0.01)Tj
1.8708 0 Td
(M)Tj
1.1112 0 Td
(EDTA,)Tj
3.0444 0 Td
(pH)Tj
1.508 0 Td
(8.0.)Tj
1.6214 0 Td
(The)Tj
1.7801 0 Td
(resulting)Tj
3.7304 0 Td
(extract)Tj
2.9367 0 Td
(was)Tj
1.6951 0 Td
(centrifugated)Tj
5.5162 0 Td
(at)Tj
0.9524 0 Td
(10,000)Tj
2.8233 0 Td
(rpm)Tj
1.9332 0 Td
(for)Tj
1.3833 0 Td
(20)Tj
-36.2208 -1.3039 Td
(min)Tj
1.8538 0 Td
(\(Beckman)Tj
4.286 0 Td
(J2-21)Tj
2.3243 0 Td
(Centrifuge,)Tj
4.7225 0 Td
(JA-14)Tj
2.5399 0 Td
(rotor\).)Tj
2.8403 0 Td
(The)Tj
1.7801 0 Td
(virus)Tj
2.1997 0 Td
(suspension)Tj
4.6431 0 Td
(was)Tj
1.7008 0 Td
(then)Tj
2.0182 0 Td
(centrifuged)Tj
4.7849 0 Td
(at)Tj
-35.6936 -1.2982 Td
(110)Tj
1.644 0 Td
(000)Tj
1.6498 0 Td
(g)Tj
0.686 0 Td
(for)Tj
1.3833 0 Td
(3)Tj
0.6973 0 Td
(h)Tj
0.7483 0 Td
(\(Hitachi)Tj
3.5433 0 Td
(Koki)Tj
2.1657 0 Td
(himac)Tj
2.6929 0 Td
(CP100WX)Tj
4.4843 0 Td
(Ultracentrifuge,)Tj
6.5707 0 Td
(P50AT2)Tj
3.4979 0 Td
(or)Tj
1.0999 0 Td
(P70AT2)Tj
3.5036 0 Td
(rotor\).)Tj
-34.367 -1.2983 Td
(The)Tj
1.7801 0 Td
(pellets)Tj
2.7382 0 Td
(were)Tj
2.1203 0 Td
(suspended)Tj
4.4391 0 Td
(overnight)Tj
4.0818 0 Td
(at)Tj
0.9525 0 Td
(4C)Tj
1.7518 0 Td
(in)Tj
1.0374 0 Td
(0.05)Tj
1.8709 0 Td
(M)Tj
1.1112 0 Td
(Tris-HC1,)Tj
4.3143 0 Td
(0.01)Tj
1.8708 0 Td
(M)Tj
1.1169 0 Td
(EDTA,)Tj
3.0444 0 Td
(pH)Tj
1.5023 0 Td
(8.0)Tj
1.4003 0 Td
(and)Tj
-35.1323 -1.3039 Td
(clarified)Tj
3.4695 0 Td
(by)Tj
1.1792 0 Td
(centrifugation)Tj
5.8904 0 Td
(at)Tj
0.9524 0 Td
(10,000)Tj
2.8233 0 Td
(g)Tj
0.6803 0 Td
(for)Tj
1.389 0 Td
(20)Tj
1.1735 0 Td
(min.)Tj
2.0807 0 Td
(Then)Tj
2.33 0 Td
(8)Tj
0.6917 0 Td
(ml)Tj
1.2869 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.474 0 Td
(virus)Tj
2.1997 0 Td
(suspension)Tj
4.6431 0 Td
(per)Tj
1.5307 0 Td
(10)Tj
1.1735 0 Td
(ml)Tj
-35.9827 -1.2983 Td
(S303922A)Tj
4.2349 0 Td
(-)Tj
0.5669 0 Td
(10PC)Tj
2.3924 0 Td
(BOTTLE)Tj
3.8778 0 Td
(\(B\))Tj
1.4797 0 Td
(tube)Tj
1.9786 0 Td
(was)Tj
1.7007 0 Td
(underlaid)Tj
4.0932 0 Td
(with)Tj
1.9956 0 Td
(2)Tj
0.6973 0 Td
(ml)Tj
1.2813 0 Td
(of)Tj
1.0148 0 Td
(30%)Tj
1.9332 0 Td
(sucrose)Tj
3.1918 0 Td
(\(w/v,)Tj
2.2563 0 Td
(in)Tj
1.0319 0 Td
(the)Tj
-33.7264 -1.2983 Td
(extraction)Tj
4.2576 0 Td
(buffer\).)Tj
3.1861 0 Td
(Samples)Tj
3.4979 0 Td
(were)Tj
2.126 0 Td
(centrifugated)Tj
5.5162 0 Td
(at)Tj
0.9524 0 Td
(110,000)Tj
3.2995 0 Td
(g)Tj
0.686 0 Td
(for)Tj
1.3833 0 Td
(4)Tj
0.6973 0 Td
(h)Tj
0.7484 0 Td
(\(Hitachi)Tj
3.5432 0 Td
(Koki)Tj
2.1657 0 Td
(himac)Tj
-32.0596 -1.3039 Td
(CP100WX)Tj
4.4843 0 Td
(Ultracentrifuge,)Tj
6.5707 0 Td
(P70AT2)Tj
3.5036 0 Td
(rotor\).)Tj
2.8403 0 Td
(The)Tj
1.7801 0 Td
(pellets)Tj
2.7383 0 Td
(were)Tj
2.1203 0 Td
(resuspended)Tj
5.2327 0 Td
(in)Tj
1.0375 0 Td
(0.025)Tj
2.3471 0 Td
(M)Tj
1.1111 0 Td
(Tris-HCl)Tj
-33.766 -1.2982 Td
(buffer,)Tj
2.8516 0 Td
(pH)Tj
1.5023 0 Td
(8.0;)Tj
1.6214 0 Td
(0.01)Tj
1.8709 0 Td
(M)Tj
1.1168 0 Td
(Tris-HCl,)Tj
4.0819 0 Td
(pH)Tj
1.5024 0 Td
(7.5)Tj
1.4003 0 Td
(or)Tj
1.0941 0 Td
(water.)Tj
2.6589 0 Td
(The)Tj
1.7858 0 Td
(resulting)Tj
3.7247 0 Td
(virus)Tj
2.2054 0 Td
(solution)Tj
3.4809 0 Td
(was)Tj
1.7008 0 Td
(centrifu-)Tj
-32.5982 -1.2983 Td
(gated)Tj
2.3697 0 Td
(at)Tj
0.9524 0 Td
(10,000)Tj
2.8233 0 Td
(g)Tj
0.6803 0 Td
(for)Tj
1.389 0 Td
(20)Tj
1.1735 0 Td
(min.)Tj
2.0807 0 Td
(Virion)Tj
2.8743 0 Td
(concentration)Tj
5.8166 0 Td
(was)Tj
1.7008 0 Td
(estimated)Tj
4.0705 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0535 0 Td
([)Tj
0.83 0.64 0.02 0 k
(1)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(10)Tj
0 g
(].)Tj
2.778 0 Td
(The)Tj
1.7858 0 Td
(viral)Tj
-35.5632 -1.3039 Td
(preparation)Tj
4.9095 0 Td
(was)Tj
1.6951 0 Td
(stored)Tj
2.7099 0 Td
(at)Tj
0.9525 0 Td
(4C)Tj
1.7518 0 Td
(or)Tj
1.0998 0 Td
(frozen)Tj
2.7723 0 Td
(at)Tj
0.9524 0 Td
(-20C)Tj
2.5739 0 Td
(for)Tj
1.3889 0 Td
(long-term)Tj
4.252 0 Td
(storage.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 498.1039 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Coat)Tj
2.3102 0 Td
(protein)Tj
3.2315 0 Td
(isolation)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 481.096 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(The)Tj
1.7801 0 Td
(method)Tj
3.3222 0 Td
(of)Tj
1.0148 0 Td
(salt)Tj
1.5647 0 Td
(deproteinization)Tj
6.8371 0 Td
(with)Tj
1.9956 0 Td
(2M)Tj
1.5874 0 Td
(LiCl)Tj
1.9389 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0535 0 Td
(previously)Tj
4.3483 0 Td
([)Tj
0.83 0.64 0.02 0 k
(10)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(24)Tj
0 g
(].)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 453.8267 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Stability)Tj
3.6756 0 Td
(analysis)Tj
3.7843 0 Td
(of)Tj
1.0441 0 Td
(AltMV)Tj
2.8677 0 Td
(virions)Tj
3.0709 0 Td
(and)Tj
1.8661 0 Td
(virus-like)Tj
4.1103 0 Td
(particles)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 436.8188 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(The)Tj
1.7801 0 Td
(AltMV)Tj
3.0387 0 Td
(virions)Tj
2.9934 0 Td
(and)Tj
1.7291 0 Td
(virus-like)Tj
3.9912 0 Td
(particles)Tj
3.5603 0 Td
(were)Tj
2.1203 0 Td
(incubated)Tj
4.1782 0 Td
(at)Tj
0.9524 0 Td
(room)Tj
2.4265 0 Td
(temperature)Tj
5.125 0 Td
(for)Tj
1.3889 0 Td
(1)Tj
0.6974 0 Td
(hour)Tj
-33.9815 -1.2982 Td
(under)Tj
2.6191 0 Td
(different)Tj
3.6567 0 Td
(conditions)Tj
4.4731 0 Td
(\(distilled)Tj
3.7247 0 Td
(water;)Tj
2.6588 0 Td
(0.15)Tj
1.8709 0 Td
(M)Tj
1.1112 0 Td
(NaCl;)Tj
2.5285 0 Td
(0.01)Tj
1.8708 0 Td
(M)Tj
1.1112 0 Td
(Tris-HCl,)Tj
4.0818 0 Td
(pH)Tj
1.5081 0 Td
(7.5;)Tj
1.6214 0 Td
(0.01)Tj
1.8708 0 Td
(M)Tj
-34.7071 -1.2983 Td
(Tris-HCl,)Tj
4.0818 0 Td
(0.15)Tj
1.8709 0 Td
(M)Tj
1.1168 0 Td
(NaCl,)Tj
2.5228 0 Td
(pH)Tj
1.5024 0 Td
(7.5;)Tj
1.6271 0 Td
(mouse)Tj
2.8573 0 Td
(serum\).)Tj
3.2881 0 Td
(Results)Tj
3.0614 0 Td
(were)Tj
2.1203 0 Td
(analyzed)Tj
3.7021 0 Td
(by)Tj
1.1792 0 Td
(electron)Tj
3.4639 0 Td
(microscopy.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 383.5842 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Electron)Tj
3.8456 0 Td
(microscopy)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 366.5763 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(The)Tj
1.7801 0 Td
(samples)Tj
3.3845 0 Td
(were)Tj
2.1203 0 Td
(prepared)Tj
3.8098 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0478 0 Td
(by)Tj
1.1849 0 Td
(Nikitin)Tj
3.1011 0 Td
(et)Tj
0.9411 0 Td
(al.)Tj
1.1225 0 Td
([)Tj
0.83 0.64 0.02 0 k
(25)Tj
0 g
(])Tj
1.8538 0 Td
(and)Tj
1.7292 0 Td
(examined)Tj
4.1329 0 Td
(either)Tj
2.5284 0 Td
(in)Tj
1.0375 0 Td
(a)Tj
0.652 0 Td
(trans-)Tj
-34.4407 -1.3039 Td
(mission)Tj
3.3505 0 Td
(electron)Tj
3.4639 0 Td
(microscope)Tj
4.8359 0 Td
(JEOL)Tj
2.4037 0 Td
(JEM-1400)Tj
4.269 0 Td
(TEM)Tj
2.2904 0 Td
(\(JEOL,)Tj
2.9707 0 Td
(Japan\))Tj
2.8232 0 Td
(operating)Tj
4.0536 0 Td
(at)Tj
0.9524 0 Td
(80)Tj
1.1679 0 Td
(kV,)Tj
1.6384 0 Td
(or)Tj
1.0998 0 Td
(JEOL)Tj
-35.3194 -1.2982 Td
(JEM-2100)Tj
4.2632 0 Td
(\(JEOL,)Tj
2.9707 0 Td
(Japan\))Tj
2.8233 0 Td
(operating)Tj
4.0535 0 Td
(at)Tj
0.9525 0 Td
(200)Tj
1.644 0 Td
(kV.)Tj
1.6385 0 Td
(Images)Tj
3.0614 0 Td
(were)Tj
2.1203 0 Td
(recorded)Tj
3.7984 0 Td
(on)Tj
1.2755 0 Td
(an)Tj
1.2019 0 Td
(Olympus)Tj
3.8948 0 Td
(Quemesa)Tj
-33.698 -1.2983 Td
(digital)Tj
2.7269 0 Td
(camera)Tj
3.1237 0 Td
(under)Tj
2.6192 0 Td
(the)Tj
1.4684 0 Td
(control)Tj
3.1237 0 Td
(of)Tj
1.0148 0 Td
(iTEM)Tj
2.5568 0 Td
(software)Tj
3.5887 0 Td
(\(Olympus)Tj
4.2349 0 Td
(Soft)Tj
1.7858 0 Td
(Imaging)Tj
3.5603 0 Td
(Solutions)Tj
3.9628 0 Td
(GmbH,)Tj
-33.766 -1.3039 Td
(Germany\).)Tj
4.5524 0 Td
(The)Tj
1.7801 0 Td
(length)Tj
2.7326 0 Td
(and)Tj
1.7291 0 Td
(width)Tj
2.5228 0 Td
(of)Tj
1.0148 0 Td
(viral)Tj
1.9956 0 Td
(and)Tj
1.7291 0 Td
(virus-like)Tj
3.9855 0 Td
(particles)Tj
3.566 0 Td
(were)Tj
2.1203 0 Td
(measured)Tj
4.1102 0 Td
(manually)Tj
-31.8385 -1.2983 Td
(from)Tj
2.2053 0 Td
(digital)Tj
2.7269 0 Td
(prints)Tj
2.5795 0 Td
(using)Tj
2.3924 0 Td
(scientific)Tj
3.7871 0 Td
(image)Tj
2.6305 0 Td
(manipulation)Tj
5.6353 0 Td
(software)Tj
3.5886 0 Td
(ImageJ)Tj
3.0274 0 Td
(\(National)Tj
4.0308 0 Td
(Institutes)Tj
3.9458 0 Td
(of)Tj
-36.5496 -1.2982 Td
(Health,)Tj
3.1407 0 Td
(USA\).)Tj
2.6816 0 Td
(To)Tj
1.3379 0 Td
(obtain)Tj
2.7836 0 Td
(the)Tj
1.4684 0 Td
(3D)Tj
1.4343 0 Td
(reconstructions,)Tj
6.6784 0 Td
(the)Tj
1.474 0 Td
(images)Tj
2.9877 0 Td
(were)Tj
2.1203 0 Td
(captured)Tj
3.7474 0 Td
(with)Tj
1.9955 0 Td
(an)Tj
1.2019 0 Td
(Ultrascan)Tj
-33.0517 -1.2983 Td
(1000XP)Tj
3.3335 0 Td
(4x4)Tj
1.6384 0 Td
(pixel)Tj
2.1373 0 Td
(CCD)Tj
2.2847 0 Td
(camera)Tj
3.1238 0 Td
(\(Gatan\))Tj
3.3278 0 Td
(at)Tj
0.9525 0 Td
(x40,000)Tj
3.2881 0 Td
(magnification)Tj
5.7883 0 Td
(with)Tj
2.0013 0 Td
(an)Tj
1.2019 0 Td
(underfocus)Tj
4.7225 0 Td
(of)Tj
1.0147 0 Td
(1.5)Tj
-34.8148 -1.3039 Td
(2.8)Tj
1.3946 0 Td
(um.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 235.2755 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Cryoelectron)Tj
5.8205 0 Td
(microscopy)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 218.2677 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(The)Tj
1.7801 0 Td
(AltMV)Tj
3.0387 0 Td
(samples)Tj
3.3789 0 Td
(were)Tj
2.126 0 Td
(diluted)Tj
3.0387 0 Td
(to)Tj
1.0261 0 Td
(a)Tj
0.652 0 Td
(final)Tj
2.0012 0 Td
(concentration)Tj
5.8224 0 Td
(of)Tj
1.0148 0 Td
(1.5mg/ml)Tj
4.0761 0 Td
(in)Tj
1.0375 0 Td
(25mM)Tj
2.88 0 Td
(Tris-HCl,)Tj
4.0875 0 Td
(pH)Tj
-35.96 -1.2982 Td
(7.5.)Tj
1.6214 0 Td
(3.5)Tj
/F11 1 Tf
1.3946 0 Td
[()]TJ
/F7 1 Tf
(l)Tj
0.9751 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(AltMV)Tj
3.0388 0 Td
(was)Tj
1.6951 0 Td
(applied)Tj
3.1521 0 Td
(to)Tj
1.0261 0 Td
(glow-discharged)Tj
6.8031 0 Td
(Quantifoil)Tj
4.3483 0 Td
(1.2/1.3)Tj
2.9084 0 Td
(grid)Tj
1.8481 0 Td
(\(Quantifoil)Tj
-31.2942 -1.2983 Td
(Micro)Tj
2.6815 0 Td
(Tools)Tj
2.4491 0 Td
(GmbH,)Tj
3.2542 0 Td
(Germany\).)Tj
4.5524 0 Td
(The)Tj
1.7858 0 Td
(grids)Tj
2.211 0 Td
(were)Tj
2.126 0 Td
(then)Tj
2.0182 0 Td
(blotted)Tj
3.0274 0 Td
(and)Tj
1.7291 0 Td
(frozen)Tj
2.7723 0 Td
(in)Tj
1.0318 0 Td
(liquid)Tj
2.5625 0 Td
(ethane)Tj
2.88 0 Td
(using)Tj
-35.0813 -1.3039 Td
(Vitrobot)Tj
3.668 0 Td
(\(Termo)Tj
3.2938 0 Td
(Fisher)Tj
2.6986 0 Td
(Scientific,)Tj
4.1272 0 Td
(USA\).)Tj
2.6815 0 Td
(Low)Tj
1.9503 0 Td
(dose)Tj
2.0409 0 Td
(images)Tj
2.9877 0 Td
(\(2025)Tj
2.982 0 Td
(e)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 468.5102 183.7417 cm
BT
/F11 1 Tf
1 TL
-0.0053 Tc
0 0 Td
[()]TJ
ET
Q
q
1 j
1 J
0 w
10 0 0 10 472.7622 179.2629 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(/)Tj
/F11 1 Tf
[()]TJ
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 482.9102 183.7417 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 486.4818 179.2629 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\))Tj
0.5612 0 Td
(were)Tj
2.1203 0 Td
(collected)Tj
3.7417 0 Td
(on)Tj
1.2756 0 Td
(a)Tj
-36.3455 -1.2982 Td
(Tecnai)Tj
2.9253 0 Td
(G12)Tj
1.8879 0 Td
(Spirit)Tj
2.4151 0 Td
(Twin)Tj
2.3301 0 Td
(electron)Tj
3.4639 0 Td
(microscope)Tj
4.8415 0 Td
(\(Termo)Tj
3.2939 0 Td
(Fisher)Tj
2.6985 0 Td
(Scientific,)Tj
4.1272 0 Td
(USA\))Tj
2.4547 0 Td
(operated)Tj
3.7247 0 Td
(at)Tj
0.9525 0 Td
(120)Tj
-35.1153 -1.2983 Td
(kV,)Tj
1.6327 0 Td
(using)Tj
2.3925 0 Td
(a)Tj
0.6519 0 Td
(4x4)Tj
1.6385 0 Td
(Eagle)Tj
2.3584 0 Td
(CCD)Tj
2.2847 0 Td
(camera)Tj
3.1237 0 Td
(\(Termo)Tj
3.2939 0 Td
(Fisher)Tj
2.6985 0 Td
(Scientific,)Tj
4.1273 0 Td
(USA\).)Tj
2.6815 0 Td
(A)Tj
0.9014 0 Td
(defocus)Tj
3.2768 0 Td
(range)Tj
2.4661 0 Td
(between)Tj
-33.5279 -1.3039 Td
(0.8)Tj
1.3946 0 Td
(and)Tj
1.7292 0 Td
(3.5)Tj
/F11 1 Tf
1.4003 0 Td
[()]TJ
/F7 1 Tf
(m)Tj
1.542 0 Td
(underfocus)Tj
4.7282 0 Td
(was)Tj
1.7007 0 Td
(used.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 112.9889 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Image)Tj
2.9716 0 Td
(processing)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 95.9811 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Image)Tj
2.6985 0 Td
(processing)Tj
4.4674 0 Td
(was)Tj
1.7008 0 Td
(carried)Tj
3.0387 0 Td
(out)Tj
1.5534 0 Td
(using)Tj
2.3867 0 Td
(the)Tj
1.4684 0 Td
(SPRING)Tj
3.668 0 Td
(software)Tj
3.5886 0 Td
([)Tj
0.83 0.64 0.02 0 k
(26)Tj
0 g
(].)Tj
2.075 0 Td
(Briefly,)Tj
3.0784 0 Td
(the)Tj
1.4683 0 Td
(images)Tj
2.9934 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
-35.2004 -1.2982 Td
(viruses)Tj
2.9876 0 Td
(and)Tj
1.7292 0 Td
(VLPs)Tj
2.3697 0 Td
(were)Tj
2.126 0 Td
(cut)Tj
1.4683 0 Td
(out)Tj
1.5534 0 Td
(into)Tj
1.8425 0 Td
(100x100)Tj
3.5376 0 Td
(pixel)Tj
2.1373 0 Td
(square)Tj
2.846 0 Td
(segments)Tj
3.9175 0 Td
(with)Tj
2.0012 0 Td
(an)Tj
1.2019 0 Td
(overlap)Tj
3.1748 0 Td
(of)Tj
1.0148 0 Td
(360)Tj
/F11 1 Tf
1.6441 0 Td
[()]TJ
/F7 1 Tf
0.907 0 Td
(in)Tj
ET
Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 107.1496 23.6976 re
W* n
q
107.0929 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
Q
Q
q
0 0 612 792 re
W* n
1 j
1 J
0 w
7.9999 0 0 7.9999 390.2173 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(Structure)Tj
4.2165 0 Td
(and)Tj
1.8567 0 Td
(properties)Tj
4.578 0 Td
(of)Tj
1.0418 0 Td
(Alternanthe)Tj
5.0244 0 Td
(ra)Tj
1.0914 0 Td
(mosaic)Tj
3.3378 0 Td
(virus)Tj
ET
Q
q
1 j
1 J
0 w
36 48.0002 m
576 48.0002 l
576 47.5002 l
36 47.5002 l
f*
0.83 0.64 0.02 0 k
81.5811 34.9228 m
237.8835 34.9228 l
h
f*
0 g
7.9999 0 0 7.9999 36 36 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(PLOS)Tj
2.8559 0 Td
(ONE)Tj
2.3669 0 Td
(|)Tj
0.83 0.64 0.02 0 k
0.4748 0 Td
(https://doi.or)Tj
5.4142 0 Td
(g/10.137)Tj
3.8198 0 Td
(1/journal.po)Tj
5.1236 0 Td
(ne.01838)Tj
4.089 0 Td
(24)Tj
0 g
2.0906 0 Td
(August)Tj
3.2882 0 Td
(24,)Tj
1.5874 0 Td
(2017)Tj
34.0302 0 Td
(3)Tj
0.7654 0 Td
(/)Tj
ET
endstream
endobj
57 0 obj
<>stream
endstream
endobj
58 0 obj
<>stream
endstream
endobj
59 0 obj
<>stream
endstream
endobj
60 0 obj
<>stream
endstream
endobj
61 0 obj
<>stream
endstream
endobj
62 0 obj
<>stream
endstream
endobj
63 0 obj
<>stream
endstream
endobj
64 0 obj
<>stream
endstream
endobj
65 0 obj
<>stream
endstream
endobj
66 0 obj
<>stream
BT
66.402 0 Td
(11)Tj
ET
Q
endstream
endobj
67 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 12 0 R/Annots 68 0 R/Contents 69 0 R/TrimBox[0 0 612 792]>>
endobj
68 0 obj
[70 0 R 71 0 R 72 0 R 73 0 R 74 0 R 75 0 R 76 0 R 77 0 R 78 0 R 79 0 R 80 0 R 81 0 R 82 0 R]
endobj
70 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref027)>>
endobj
71 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref028)>>
endobj
72 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref029)>>
endobj
73 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref030)>>
endobj
74 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref031)>>
endobj
75 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref001)>>
endobj
76 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref005)>>
endobj
77 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref010)>>
endobj
78 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref015)>>
endobj
79 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref001)>>
endobj
80 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref032)>>
endobj
81 0 obj
<>/Border[0 0 0]/Dest(Rpone.0183824.ref033)>>
endobj
82 0 obj
<>/Border[0 0 0]/A 83 0 R>>
endobj
83 0 obj
<>
endobj
69 0 obj
[84 0 R 85 0 R 86 0 R 87 0 R 88 0 R 89 0 R 90 0 R 91 0 R 92 0 R 93 0 R 94 0 R 95 0 R 96 0 R 97 0 R 98 0 R]
endobj
84 0 obj
<>stream
q
0.83 0.64 0.02 0 k
328.6488 706.337 m
338.1732 706.337 l
h
f*
359.0929 693.2976 m
368.5606 693.2976 l
h
f*
442.148 446.3433 m
451.6724 446.3433 l
h
f*
546.8598 364.1386 m
556.3843 364.1386 l
h
f*
225.3543 237.9402 m
234.8787 237.9402 l
h
f*
203.4142 198.9354 m
208.1764 198.9354 l
h
f*
314.7591 198.9354 m
319.5213 198.9354 l
h
f*
431.7165 198.9354 m
441.1843 198.9354 l
h
f*
443.452 198.9354 m
452.9764 198.9354 l
h
f*
203.4142 159.9307 m
208.1764 159.9307 l
h
f*
307.0488 107.9433 m
316.5732 107.9433 l
h
f*
485.178 94.9039 m
494.7024 94.9039 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 200.0125 707.4141 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(the)Tj
1.4683 0 Td
(EMAN)Tj
/F8 1 Tf
3.1011 0 Td
[()6()15()12()4()19()-4()2()]TJ
/F7 1 Tf
4.2462 0 Td
(program)Tj
3.7077 0 Td
([)Tj
0.83 0.64 0.02 0 k
(27)Tj
0 g
(])Tj
1.8482 0 Td
(and)Tj
1.7348 0 Td
(appended)Tj
4.1386 0 Td
(in)Tj
1.0374 0 Td
(a)Tj
0.652 0 Td
(single)Tj
2.5285 0 Td
(stack.)Tj
2.4604 0 Td
(The)Tj
1.7859 0 Td
(CTF)Tj
2.0182 0 Td
(was)Tj
1.7008 0 Td
(determined)Tj
-32.4281 -1.3039 Td
(for)Tj
1.3832 0 Td
(each)Tj
2.0296 0 Td
(micrograph)Tj
4.9266 0 Td
(using)Tj
2.3868 0 Td
(CTFFIND3)Tj
4.8415 0 Td
([)Tj
0.83 0.64 0.02 0 k
(28)Tj
0 g
(])Tj
1.8482 0 Td
(incorporated)Tj
5.4198 0 Td
(into)Tj
1.8369 0 Td
(the)Tj
1.474 0 Td
(SPRING)Tj
3.668 0 Td
(program.)Tj
3.9288 0 Td
(Thereby)Tj
-33.7434 -1.2982 Td
(1180)Tj
2.1203 0 Td
(single)Tj
2.5341 0 Td
(frames)Tj
2.914 0 Td
(were)Tj
2.126 0 Td
(collected)Tj
3.736 0 Td
(for)Tj
1.389 0 Td
(AltMV)Tj
3.033 0 Td
(VLPs,)Tj
2.6022 0 Td
(and)Tj
1.7291 0 Td
(230)Tj
1.6441 0 Td
(for)Tj
1.389 0 Td
(AltMV)Tj
3.0331 0 Td
(virions.)Tj
3.2258 0 Td
(Then,)Tj
2.5568 0 Td
(all)Tj
1.1452 0 Td
(the)Tj
-35.1777 -1.2983 Td
(AltMV)Tj
3.033 0 Td
(virion)Tj
2.6362 0 Td
(particles)Tj
3.566 0 Td
(were)Tj
2.1203 0 Td
(classified)Tj
3.821 0 Td
(into)Tj
1.8369 0 Td
(10)Tj
1.1735 0 Td
(classes)Tj
2.829 0 Td
(with)Tj
1.9956 0 Td
(~22)Tj
1.7518 0 Td
(particles)Tj
3.5659 0 Td
(per)Tj
1.5307 0 Td
(class.)Tj
2.2677 0 Td
(AltMV)Tj
3.0387 0 Td
(VLPs)Tj
-35.1663 -1.2983 Td
(were)Tj
2.1203 0 Td
(classified)Tj
3.821 0 Td
(into)Tj
1.8369 0 Td
(50)Tj
1.1735 0 Td
(classes)Tj
2.829 0 Td
(with)Tj
2.0012 0 Td
(~22)Tj
1.7461 0 Td
(particles)Tj
3.566 0 Td
(per)Tj
1.5307 0 Td
(class.)Tj
2.2734 0 Td
(For)Tj
1.6214 0 Td
(all)Tj
1.1508 0 Td
(class)Tj
2.041 0 Td
(sums,)Tj
2.5171 0 Td
(the)Tj
1.4684 0 Td
(Fourier)Tj
3.2144 0 Td
(trans-)Tj
-34.9112 -1.3039 Td
(form)Tj
2.2053 0 Td
(was)Tj
1.7008 0 Td
(carried)Tj
3.0387 0 Td
(out.)Tj
-5.7486 -1.2982 Td
(To)Tj
1.3379 0 Td
(determine)Tj
4.3256 0 Td
(the)Tj
1.4684 0 Td
(parameters)Tj
4.6771 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(helix,)Tj
2.3754 0 Td
(three-dimensional)Tj
7.5628 0 Td
(reconstructions)Tj
6.4572 0 Td
(with)Tj
2.0013 0 Td
(low)Tj
1.6554 0 Td
(reso-)Tj
-35.5405 -1.2983 Td
(lution)Tj
2.6135 0 Td
(were)Tj
2.1203 0 Td
(constructed)Tj
4.9322 0 Td
(at)Tj
0.9525 0 Td
(various)Tj
3.1407 0 Td
(parameters)Tj
4.6772 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(helix)Tj
2.1487 0 Td
(angle)Tj
2.3414 0 Td
(\(from)Tj
2.5454 0 Td
(34)Tj
1.1679 0 Td
(to)Tj
1.0262 0 Td
(44)Tj
/F11 1 Tf
1.1735 0 Td
[()]TJ
/F7 1 Tf
0.9014 0 Td
(in)Tj
1.0375 0 Td
(0.1)Tj
/F11 1 Tf
1.3946 0 Td
[()]TJ
/F7 1 Tf
-34.6561 -1.3039 Td
(steps\))Tj
2.5171 0 Td
(and)Tj
1.7348 0 Td
(the)Tj
1.4683 0 Td
(number)Tj
3.4186 0 Td
(of)Tj
1.0148 0 Td
(subunits)Tj
3.617 0 Td
(\(from)Tj
2.5455 0 Td
(8)Tj
0.6973 0 Td
(to)Tj
1.0261 0 Td
(10)Tj
/F11 1 Tf
1.1679 0 Td
[()]TJ
/F7 1 Tf
0.9071 0 Td
(in)Tj
1.0318 0 Td
(increments)Tj
4.7055 0 Td
(of)Tj
1.0148 0 Td
(0.01)Tj
/F11 1 Tf
1.8765 0 Td
[()]TJ
/F7 1 Tf
(\))Tj
1.2415 0 Td
(per)Tj
1.5307 0 Td
(rotation)Tj
3.4526 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
-35.9827 -1.2983 Td
(helix.)Tj
2.3754 0 Td
(As)Tj
1.2642 0 Td
(a)Tj
0.652 0 Td
(result,)Tj
2.6759 0 Td
(50)Tj
1.1678 0 Td
(reconstructions)Tj
6.4573 0 Td
(were)Tj
2.126 0 Td
(generated.)Tj
4.3596 0 Td
(The)Tj
1.7802 0 Td
(pitch)Tj
2.2564 0 Td
(of)Tj
1.0147 0 Td
(the)Tj
1.4684 0 Td
(helix)Tj
2.1543 0 Td
(was)Tj
1.6951 0 Td
(estimated)Tj
-31.4473 -1.2983 Td
(using)Tj
2.3867 0 Td
(50)Tj
1.1735 0 Td
(best)Tj
1.8085 0 Td
(reconstructions)Tj
6.4573 0 Td
(based)Tj
2.4718 0 Td
(on)Tj
1.2756 0 Td
(the)Tj
1.474 0 Td
(average)Tj
3.2258 0 Td
(cross-correlation)Tj
6.9902 0 Td
(index)Tj
2.4491 0 Td
(and)Tj
1.7348 0 Td
(amounted)Tj
4.303 0 Td
(to)Tj
-35.7503 -1.3039 Td
(35.7)Tj
/F11 1 Tf
1.8708 0 Td
[()]TJ
/F7 1 Tf
(.)Tj
1.1282 0 Td
(The)Tj
1.7858 0 Td
(determination)Tj
5.9527 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(number)Tj
3.4185 0 Td
(of)Tj
1.0148 0 Td
(subunits)Tj
3.617 0 Td
(per)Tj
1.5364 0 Td
(helix)Tj
2.1486 0 Td
(is)Tj
0.8447 0 Td
(based)Tj
2.4775 0 Td
(on)Tj
1.2756 0 Td
(an)Tj
1.2019 0 Td
(analysis)Tj
3.3221 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
-35.0926 -1.2982 Td
(amplitude)Tj
4.2802 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(correlation)Tj
4.6204 0 Td
(between)Tj
3.5263 0 Td
(the)Tj
1.4683 0 Td
(class-average)Tj
5.4028 0 Td
(and)Tj
1.7292 0 Td
(the)Tj
1.474 0 Td
(re-projections)Tj
5.845 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(3D)Tj
1.4343 0 Td
(mod-)Tj
-34.7468 -1.2983 Td
(els.)Tj
1.474 0 Td
(The)Tj
1.7858 0 Td
(subsequent)Tj
4.6998 0 Td
(improvement)Tj
5.6749 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(reconstruction)Tj
6.0945 0 Td
(was)Tj
1.7008 0 Td
(carried)Tj
3.0387 0 Td
(out)Tj
1.5534 0 Td
(in)Tj
1.0374 0 Td
(50)Tj
1.1679 0 Td
(iterations,)Tj
4.2236 0 Td
(until)Tj
-34.9339 -1.3039 Td
(the)Tj
1.4683 0 Td
(resolution)Tj
4.2803 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(electron)Tj
3.4583 0 Td
(density)Tj
3.101 0 Td
(map)Tj
1.9956 0 Td
(did)Tj
1.5421 0 Td
(not)Tj
1.5703 0 Td
(cease)Tj
2.2847 0 Td
(to)Tj
1.0262 0 Td
(improve.)Tj
-22.0137 -1.2983 Td
(The)Tj
1.7744 0 Td
(resolution)Tj
4.2747 0 Td
(of)Tj
1.0034 0 Td
(the)Tj
1.4627 0 Td
(final)Tj
1.9956 0 Td
(3D)Tj
1.4229 0 Td
(reconstructions)Tj
6.4517 0 Td
(was)Tj
1.6894 0 Td
(estimated)Tj
4.0649 0 Td
(by)Tj
1.1735 0 Td
(the)Tj
1.4627 0 Td
(Fourier)Tj
3.2088 0 Td
(shell)Tj
2.0182 0 Td
(correlation)Tj
-33.1991 -1.2982 Td
(\(FSC\))Tj
2.5681 0 Td
(using)Tj
2.3868 0 Td
(a)Tj
0.6519 0 Td
(0.143)Tj
2.3528 0 Td
(criterion.)Tj
3.9231 0 Td
(This)Tj
1.9842 0 Td
(was)Tj
1.7008 0 Td
(done)Tj
2.2224 0 Td
(by)Tj
1.1848 0 Td
(splitting)Tj
3.4866 0 Td
(the)Tj
1.474 0 Td
(final)Tj
2.0013 0 Td
(average)Tj
3.2258 0 Td
(and)Tj
1.7291 0 Td
(symmetry)Tj
-30.8917 -1.2983 Td
(related)Tj
2.9423 0 Td
(views)Tj
2.4094 0 Td
(into)Tj
1.8425 0 Td
(even)Tj
2.0693 0 Td
(and)Tj
1.7291 0 Td
(odd)Tj
1.7859 0 Td
(numbers.)Tj
4.0024 0 Td
(The)Tj
1.7859 0 Td
(reconstructions)Tj
6.4572 0 Td
(from)Tj
2.2054 0 Td
(the)Tj
1.4683 0 Td
(two)Tj
1.7065 0 Td
(halves)Tj
2.6702 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(data)Tj
-35.5575 -1.3039 Td
(were)Tj
2.1203 0 Td
(used)Tj
2.0636 0 Td
(in)Tj
1.0318 0 Td
(the)Tj
1.4683 0 Td
(FSC)Tj
1.8879 0 Td
(calculation.)Tj
-7.3757 -1.2983 Td
(To)Tj
1.3379 0 Td
(generate)Tj
3.6113 0 Td
(unbiased)Tj
3.8154 0 Td
(difference)Tj
4.2066 0 Td
(maps,)Tj
2.5739 0 Td
(the)Tj
1.474 0 Td
(3D)Tj
1.4286 0 Td
(reconstructions)Tj
6.4573 0 Td
(of)Tj
1.0148 0 Td
(AltMV)Tj
3.0387 0 Td
(virions)Tj
2.9934 0 Td
(and)Tj
1.7291 0 Td
(VLPs)Tj
-34.8772 -1.2983 Td
(were)Tj
2.1203 0 Td
(aligned)Tj
3.135 0 Td
(against)Tj
3.0274 0 Td
(their)Tj
2.109 0 Td
(symmetry)Tj
4.2236 0 Td
(axis)Tj
1.7461 0 Td
(using)Tj
2.3868 0 Td
(Chimera)Tj
3.7303 0 Td
(1.9)Tj
1.3947 0 Td
([)Tj
0.83 0.64 0.02 0 k
(29)Tj
0 g
(])Tj
1.8482 0 Td
(before)Tj
2.7439 0 Td
(subtraction)Tj
4.7622 0 Td
(using)Tj
2.3924 0 Td
(the)Tj
-35.6199 -1.3039 Td
(command)Tj
4.2972 0 Td
(vop)Tj
1.7008 0 Td
(substract.)Tj
4.0252 0 Td
(To)Tj
1.3379 0 Td
(visualize)Tj
3.617 0 Td
(differences)Tj
4.5638 0 Td
(between)Tj
3.5319 0 Td
(the)Tj
1.4684 0 Td
(AltMV)Tj
3.033 0 Td
(virions)Tj
2.999 0 Td
(and)Tj
1.7292 0 Td
(VLPs)Tj
2.3754 0 Td
(the)Tj
-34.6788 -1.2982 Td
(obtained)Tj
3.7303 0 Td
(difference)Tj
4.2066 0 Td
(map)Tj
1.9899 0 Td
(was)Tj
1.7008 0 Td
(fitted)Tj
2.3244 0 Td
(into)Tj
1.8368 0 Td
(the)Tj
1.4684 0 Td
(3D)Tj
1.4343 0 Td
(reconstruction)Tj
6.0944 0 Td
(of)Tj
1.0148 0 Td
(VLP)Tj
2.0126 0 Td
(in)Tj
1.0375 0 Td
(Chimera)Tj
3.7247 0 Td
(1.9.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 395.2062 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Trypsin)Tj
3.463 0 Td
(test)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 378.1984 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(The)Tj
1.7801 0 Td
(trypsin)Tj
3.0387 0 Td
(\(Promega,)Tj
4.3653 0 Td
(10)Tj
1.1736 0 Td
(ng)Tj
1.2302 0 Td
(per)Tj
1.5364 0 Td
(1)Tj
0.6916 0 Td
(mg)Tj
1.508 0 Td
(of)Tj
1.0148 0 Td
(virions,)Tj
3.2202 0 Td
(VLPs)Tj
2.3754 0 Td
(or)Tj
1.0942 0 Td
(CP\))Tj
1.7858 0 Td
(was)Tj
1.6951 0 Td
(added)Tj
2.6645 0 Td
(to)Tj
1.0262 0 Td
(the)Tj
1.4683 0 Td
(samples,)Tj
3.6057 0 Td
(and)Tj
-35.2741 -1.2982 Td
(they)Tj
1.9218 0 Td
(were)Tj
2.126 0 Td
(incubated)Tj
4.1726 0 Td
(in)Tj
1.0374 0 Td
(100)Tj
1.6498 0 Td
(mM)Tj
1.9275 0 Td
(Tris-HCl,)Tj
4.0876 0 Td
(pH)Tj
1.5023 0 Td
(8.0)Tj
1.4003 0 Td
(for)Tj
1.3833 0 Td
(one)Tj
1.7008 0 Td
(hour)Tj
2.1543 0 Td
(at)Tj
0.9525 0 Td
(37C)Tj
2.2223 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0535 0 Td
(in)Tj
1.0375 0 Td
([)Tj
0.83 0.64 0.02 0 k
(30)Tj
0 g
(].)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 339.0236 cm
BT
/F2 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Results)Tj
3.7984 0 Td
(and)Tj
1.989 0 Td
(discussion)Tj
/F0 1 Tf
-5.7874 -1.5023 Td
(Optimization)Tj
5.7165 0 Td
(of)Tj
1.0441 0 Td
(the)Tj
1.5969 0 Td
(AltMV)Tj
2.8677 0 Td
(purification)Tj
4.9891 0 Td
(protocol)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 303.9874 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(For)Tj
1.627 0 Td
(the)Tj
1.4684 0 Td
(successful)Tj
4.1612 0 Td
(use)Tj
1.5307 0 Td
(of)Tj
1.0148 0 Td
(plant)Tj
2.2677 0 Td
(viruses)Tj
2.9877 0 Td
(in)Tj
1.0318 0 Td
(biotechnologies,)Tj
6.7237 0 Td
(it)Tj
0.7881 0 Td
(is)Tj
0.8503 0 Td
(necessary)Tj
4.0196 0 Td
(to)Tj
1.0261 0 Td
(provide)Tj
3.2882 0 Td
(the)Tj
1.4683 0 Td
(effective)Tj
-34.2536 -1.2982 Td
(accumulation,)Tj
5.9016 0 Td
(isolation)Tj
3.6567 0 Td
(and)Tj
1.7291 0 Td
(purification)Tj
4.938 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(viral)Tj
1.9956 0 Td
(particles.)Tj
3.787 0 Td
(The)Tj
1.7802 0 Td
(high)Tj
2.0069 0 Td
(yield)Tj
2.143 0 Td
(of)Tj
1.0148 0 Td
(virus)Tj
2.1996 0 Td
(per)Tj
1.5307 0 Td
(unit)Tj
-35.1663 -1.2983 Td
(of)Tj
1.0147 0 Td
(plant)Tj
2.2621 0 Td
(biomass)Tj
3.4752 0 Td
(is)Tj
0.8504 0 Td
(important)Tj
4.2803 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(economic)Tj
4.1385 0 Td
(expediency)Tj
4.6999 0 Td
(of)Tj
1.0147 0 Td
(plant)Tj
2.2621 0 Td
(viruses')Tj
3.2031 0 Td
(usage.)Tj
2.6532 0 Td
(Undoubt-)Tj
-32.3487 -1.3039 Td
(edly,)Tj
2.0976 0 Td
(among)Tj
2.9933 0 Td
(plant)Tj
2.2621 0 Td
(viruses)Tj
2.9877 0 Td
(actively)Tj
3.2201 0 Td
(used)Tj
2.0636 0 Td
(in)Tj
1.0375 0 Td
(biotechnology,)Tj
6.1228 0 Td
(the)Tj
1.474 0 Td
(Tobacco)Tj
3.6283 0 Td
(mosaic)Tj
3.0331 0 Td
(virus)Tj
2.2053 0 Td
(remains)Tj
-33.1254 -1.2983 Td
(the)Tj
1.4683 0 Td
("champion")Tj
4.9039 0 Td
(in)Tj
1.0375 0 Td
(accumulation)Tj
5.6749 0 Td
(in)Tj
1.0375 0 Td
(plants:)Tj
2.8516 0 Td
(its)Tj
1.1452 0 Td
(yield)Tj
2.1373 0 Td
(reaches)Tj
3.1861 0 Td
(up)Tj
1.2643 0 Td
(to)Tj
1.0261 0 Td
(10)Tj
1.1679 0 Td
(g)Tj
0.6859 0 Td
(/)Tj
0.55 0 Td
(1)Tj
0.6973 0 Td
(kg)Tj
1.1792 0 Td
(of)Tj
1.0148 0 Td
(green)Tj
2.4548 0 Td
(plant)Tj
2.262 0 Td
(bio-)Tj
-35.7446 -1.2982 Td
(mass)Tj
2.194 0 Td
([)Tj
0.83 0.64 0.02 0 k
(31)Tj
0 g
(].)Tj
-0.9978 -1.3039 Td
(The)Tj
1.7858 0 Td
(first)Tj
1.8028 0 Td
(stage)Tj
2.2053 0 Td
(in)Tj
1.0318 0 Td
(this)Tj
1.6781 0 Td
(work)Tj
2.2791 0 Td
(was)Tj
1.6951 0 Td
(the)Tj
1.474 0 Td
(optimization)Tj
5.3631 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(AltMV)Tj
3.0387 0 Td
(purification)Tj
4.9322 0 Td
(protocol)Tj
3.6 0 Td
(to)Tj
1.0261 0 Td
(pro-)Tj
-35.5915 -1.2983 Td
(duce)Tj
2.1203 0 Td
(large)Tj
2.1599 0 Td
(quantities)Tj
4.1556 0 Td
(of)Tj
1.0148 0 Td
(virus.)Tj
2.4264 0 Td
(Earlier,)Tj
3.1295 0 Td
(AltMV)Tj
3.033 0 Td
(was)Tj
1.7008 0 Td
(isolated)Tj
3.2825 0 Td
(from)Tj
/F8 1 Tf
2.2053 0 Td
[()1()6()2()6()11(\r)4()4()12()12( )12()]TJ
5.743 0 Td
[()16()6()16()20()16()22()7()4()7()10()4()11()]TJ
/F7 1 Tf
-30.9711 -1.2983 Td
([)Tj
0.83 0.64 0.02 0 k
(1)Tj
0 g
(],)Tj
/F8 1 Tf
1.5987 0 Td
[()35()4()7()12()7()4()16()6()]TJ
4.0818 0 Td
[()4()2()22()10()6()16()18()4()16()6()]TJ
/F7 1 Tf
5.4539 0 Td
([)Tj
0.83 0.64 0.02 0 k
(5)Tj
0 g
(])Tj
1.3776 0 Td
(and)Tj
/F8 1 Tf
1.7291 0 Td
[()23()11()-19()7( )4()4()6()7()]TJ
4.0025 0 Td
[()-8()20()16()6()12()12()-36()4()11()20()]TJ
/F7 1 Tf
4.5864 0 Td
([)Tj
0.83 0.64 0.02 0 k
(10)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(15)Tj
0 g
(].)Tj
3.2485 0 Td
(In)Tj
1.1112 0 Td
(our)Tj
1.6271 0 Td
(present)Tj
3.1634 0 Td
(work,)Tj
2.5058 0 Td
(to)Tj
-34.486 -1.3039 Td
(increase)Tj
3.4695 0 Td
(the)Tj
1.4684 0 Td
(AltMV)Tj
3.0387 0 Td
(yield,)Tj
2.3641 0 Td
(the)Tj
1.4683 0 Td
(previous)Tj
3.651 0 Td
(method)Tj
3.3222 0 Td
(of)Tj
1.0148 0 Td
(virus)Tj
2.2053 0 Td
(isolation)Tj
3.6567 0 Td
(and)Tj
1.7291 0 Td
(purification)Tj
4.9379 0 Td
(has)Tj
1.5421 0 Td
(been)Tj
-33.8681 -1.2983 Td
(improved)Tj
4.0988 0 Td
(and)Tj
1.7178 0 Td
(adapted)Tj
3.3732 0 Td
(for)Tj
1.3776 0 Td
(two)Tj
1.7008 0 Td
(plants:)Tj
/F8 1 Tf
2.8403 0 Td
[()]TJ
/F7 1 Tf
(.)Tj
/F8 1 Tf
0.9865 0 Td
[()-8()20()16()6()12()12()-36()4()11()20()]TJ
/F7 1 Tf
4.5751 0 Td
(and)Tj
/F8 1 Tf
1.7177 0 Td
[()]TJ
/F7 1 Tf
(.)Tj
/F8 1 Tf
1.1339 0 Td
[()4()2()22()10()6()16()18()4()16()6()]TJ
/F7 1 Tf
(.)Tj
5.6692 0 Td
(Geering)Tj
3.4243 0 Td
(and)Tj
1.7178 0 Td
(Thomas)Tj
-34.333 -1.2982 Td
([)Tj
0.83 0.64 0.02 0 k
(1)Tj
0 g
(])Tj
1.3719 0 Td
(propagated)Tj
4.7168 0 Td
(AltMV)Tj
3.0388 0 Td
(in)Tj
/F8 1 Tf
1.0318 0 Td
[()]TJ
/F7 1 Tf
(.)Tj
/F8 1 Tf
1.0714 0 Td
[()16()6()16()20()16()22()7()4()7()10()4()11()]TJ
/F7 1 Tf
(,)Tj
6.0321 0 Td
(yielding)Tj
3.4186 0 Td
(a)Tj
0.6576 0 Td
(23.4)Tj
1.8709 0 Td
(mg)Tj
1.5023 0 Td
(of)Tj
1.0148 0 Td
(virus)Tj
2.2054 0 Td
(per)Tj
1.5307 0 Td
(100)Tj
1.6441 0 Td
(g)Tj
0.6859 0 Td
(of)Tj
1.0148 0 Td
(infected)Tj
-32.8079 -1.2983 Td
(leaves.)Tj
2.7892 0 Td
(This)Tj
1.9843 0 Td
(is)Tj
0.8504 0 Td
(a)Tj
0.6519 0 Td
(quite)Tj
2.245 0 Td
(high)Tj
2.007 0 Td
(yield,)Tj
2.364 0 Td
(but)Tj
1.5534 0 Td
(the)Tj
1.4684 0 Td
(host)Tj
1.9162 0 Td
(plant)Tj
2.262 0 Td
(used)Tj
2.0636 0 Td
(for)Tj
1.3833 0 Td
(this)Tj
1.6724 0 Td
(cannot)Tj
2.9821 0 Td
(be)Tj
1.1508 0 Td
(considered)Tj
4.5978 0 Td
(optimal)Tj
-33.9418 -1.3039 Td
(for)Tj
1.3832 0 Td
(the)Tj
1.4741 0 Td
(accumulation)Tj
5.6749 0 Td
(of)Tj
1.0148 0 Td
(AltMV.)Tj
/F8 1 Tf
3.2598 0 Td
[()]TJ
/F7 1 Tf
(.)Tj
/F8 1 Tf
1.0658 0 Td
[()16()6()16()20()16()22()7()4()7()10()4()11()]TJ
/F7 1 Tf
5.811 0 Td
(is)Tj
0.8504 0 Td
(the)Tj
1.4683 0 Td
(host)Tj
1.9106 0 Td
(for)Tj
1.3889 0 Td
(a)Tj
0.652 0 Td
(wide)Tj
2.126 0 Td
(variety)Tj
2.9196 0 Td
(of)Tj
1.0148 0 Td
(plant)Tj
2.2677 0 Td
(viruses,)Tj
-34.2819 -1.2983 Td
(including)Tj
4.0421 0 Td
(the)Tj
1.4684 0 Td
(representatives)Tj
6.1965 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
/F8 1 Tf
1.4683 0 Td
[()23()12()11()1()-13()12()-9( )-4(\n)]TJ
/F7 1 Tf
4.3086 0 Td
(genus)Tj
2.5455 0 Td
(that)Tj
1.7802 0 Td
(can)Tj
1.627 0 Td
(lead)Tj
1.8482 0 Td
(to)Tj
1.0262 0 Td
(contamination)Tj
6.1228 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
-34.4634 -1.2982 Td
(AltMV)Tj
3.033 0 Td
(with)Tj
2.0012 0 Td
(other)Tj
2.3471 0 Td
(viruses)Tj
2.9877 0 Td
([)Tj
0.83 0.64 0.02 0 k
(32)Tj
0 g
(].)Tj
2.075 0 Td
(Therefore,)Tj
4.3936 0 Td
(we)Tj
1.3266 0 Td
(decided)Tj
3.3392 0 Td
(to)Tj
1.0262 0 Td
(use)Tj
1.5307 0 Td
(plants)Tj
2.6248 0 Td
(of)Tj
1.0148 0 Td
(portulaca)Tj
3.9799 0 Td
(and)Tj
1.7291 0 Td
(tobacco,)Tj
-33.4089 -1.3039 Td
(since)Tj
/F8 1 Tf
2.245 0 Td
[()]TJ
/F7 1 Tf
(.)Tj
/F8 1 Tf
0.9921 0 Td
[()-8()20()16()6()12()12()-36()4()11()20()]TJ
/F7 1 Tf
4.5864 0 Td
(is)Tj
0.8447 0 Td
(hardly)Tj
2.7836 0 Td
(susceptible)Tj
4.5751 0 Td
(to)Tj
1.0262 0 Td
(infection)Tj
3.7927 0 Td
(with)Tj
1.9956 0 Td
(other)Tj
2.3471 0 Td
(viruses)Tj
2.9877 0 Td
([)Tj
0.83 0.64 0.02 0 k
(33)Tj
0 g
(],)Tj
2.0749 0 Td
(and)Tj
/F8 1 Tf
1.7348 0 Td
[()]TJ
/F7 1 Tf
(.)Tj
/F8 1 Tf
1.1395 0 Td
[()4()2()22()10()6()16()18()]TJ
-33.1254 -1.2983 Td
[()16()6()]TJ
/F7 1 Tf
1.7177 0 Td
(is)Tj
0.8504 0 Td
(a)Tj
0.652 0 Td
(classical)Tj
3.4185 0 Td
(laboratory)Tj
4.3484 0 Td
(plant)Tj
2.262 0 Td
(very)Tj
1.9275 0 Td
(convenient)Tj
4.6658 0 Td
(for)Tj
1.3833 0 Td
(work.)Tj
2.5059 0 Td
(We)Tj
1.61 0 Td
(conducted)Tj
4.4277 0 Td
(a)Tj
0.652 0 Td
(passage)Tj
3.2144 0 Td
(of)Tj
ET
Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 107.1496 23.6976 re
W* n
q
107.0929 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
Q
Q
q
0 0 612 792 re
W* n
1 j
1 J
0 w
7.9999 0 0 7.9999 390.2173 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(Structure)Tj
4.2165 0 Td
(and)Tj
1.8567 0 Td
(properties)Tj
4.578 0 Td
(of)Tj
1.0418 0 Td
(Alternanthe)Tj
5.0244 0 Td
(ra)Tj
1.0914 0 Td
(mosaic)Tj
3.3378 0 Td
(virus)Tj
ET
Q
q
1 j
1 J
0 w
36 48.0002 m
576 48.0002 l
576 47.5002 l
36 47.5002 l
f*
0.83 0.64 0.02 0 k
81.5811 34.9228 m
237.8835 34.9228 l
h
f*
0 g
7.9999 0 0 7.9999 36 36 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(PLOS)Tj
2.8559 0 Td
(ONE)Tj
2.3669 0 Td
(|)Tj
0.83 0.64 0.02 0 k
0.4748 0 Td
(https://doi.or)Tj
5.4142 0 Td
(g/10.137)Tj
3.8198 0 Td
(1/journal.po)Tj
5.1236 0 Td
(ne.01838)Tj
4.089 0 Td
(24)Tj
0 g
2.0906 0 Td
(August)Tj
3.2882 0 Td
(24,)Tj
1.5874 0 Td
(2017)Tj
34.0302 0 Td
(4)Tj
0.7654 0 Td
(/)Tj
ET
endstream
endobj
85 0 obj
<>stream
endstream
endobj
86 0 obj
<>stream
endstream
endobj
87 0 obj
<>stream
endstream
endobj
88 0 obj
<>stream
endstream
endobj
89 0 obj
<>stream
endstream
endobj
90 0 obj
<>stream
endstream
endobj
91 0 obj
<>stream
endstream
endobj
92 0 obj
<>stream
endstream
endobj
93 0 obj
<>stream
endstream
endobj
94 0 obj
<>stream
endstream
endobj
95 0 obj
<>stream
endstream
endobj
96 0 obj
<>stream
endstream
endobj
97 0 obj
<>stream
endstream
endobj
98 0 obj
<>stream
BT
66.402 0 Td
(11)Tj
ET
Q
endstream
endobj
99 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 12 0 R/Annots 102 0 R/Contents 103 0 R/TrimBox[0 0 612 792]>>
endobj
100 0 obj
<>
endobj
104 0 obj
<>
endobj
105 0 obj
<>stream
%!PS-AdobeFont-1.0
%%This font is a conversion from Helvetica-BoldOblique with the following copyright notice:
%Copyright (c) 1985, 1987, 1989, 1990 Adobe Systems Incorporated. All Rights Reserved.Helvetica is a trademark of Linotype AG and/or its subsidiaries.
11 dict begin
/FontName /Helvetica-BoldOblique def
/FontType 1 def
/FontBBox {-166 -218 1114 770} readonly def
/FontMatrix [0.001 0 0 0.001 0 0] readonly def
/PaintType 0 def
/FontInfo 8 dict dup begin
/FullName (Helvetica-BoldOblique) readonly def
/BaseFontName (Helvetica-BoldOblique) def
/Notice (Copyright \(c\) 1985, 1987, 1989, 1990 Adobe Systems Incorporated. All Rights Reserved.Helvetica is a trademark of Linotype AG and/or its subsidiaries.) readonly def
end def
/Encoding 256 array
0 1 255 {1 index exch /.notdef put} for
dup 32 /space put
dup 33 /exclam put
dup 34 /quotedbl put
dup 35 /numbersign put
dup 36 /dollar put
dup 37 /percent put
dup 38 /ampersand put
dup 39 /quoteright put
dup 40 /parenleft put
dup 41 /parenright put
dup 42 /asterisk put
dup 43 /plus put
dup 44 /comma put
dup 45 /hyphen put
dup 46 /period put
dup 47 /slash put
dup 48 /zero put
dup 49 /one put
dup 50 /two put
dup 51 /three put
dup 52 /four put
dup 53 /five put
dup 54 /six put
dup 55 /seven put
dup 56 /eight put
dup 57 /nine put
dup 58 /colon put
dup 59 /semicolon put
dup 60 /less put
dup 61 /equal put
dup 62 /greater put
dup 63 /question put
dup 64 /at put
dup 65 /A put
dup 66 /B put
dup 67 /C put
dup 68 /D put
dup 69 /E put
dup 70 /F put
dup 71 /G put
dup 72 /H put
dup 73 /I put
dup 74 /J put
dup 75 /K put
dup 76 /L put
dup 77 /M put
dup 78 /N put
dup 79 /O put
dup 80 /P put
dup 81 /Q put
dup 82 /R put
dup 83 /S put
dup 84 /T put
dup 85 /U put
dup 86 /V put
dup 87 /W put
dup 88 /X put
dup 89 /Y put
dup 90 /Z put
dup 91 /bracketleft put
dup 92 /backslash put
dup 93 /bracketright put
dup 94 /asciicircum put
dup 95 /underscore put
dup 96 /quoteleft put
dup 97 /a put
dup 98 /b put
dup 99 /c put
dup 100 /d put
dup 101 /e put
dup 102 /f put
dup 103 /g put
dup 104 /h put
dup 105 /i put
dup 106 /j put
dup 107 /k put
dup 108 /l put
dup 109 /m put
dup 110 /n put
dup 111 /o put
dup 112 /p put
dup 113 /q put
dup 114 /r put
dup 115 /s put
dup 116 /t put
dup 117 /u put
dup 118 /v put
dup 119 /w put
dup 120 /x put
dup 121 /y put
dup 122 /z put
dup 123 /braceleft put
dup 124 /bar put
dup 125 /braceright put
dup 126 /asciitilde put
dup 161 /exclamdown put
dup 162 /cent put
dup 163 /sterling put
dup 164 /fraction put
dup 165 /yen put
dup 166 /florin put
dup 167 /section put
dup 168 /currency put
dup 169 /quotesingle put
dup 170 /quotedblleft put
dup 171 /guillemotleft put
dup 172 /guilsinglleft put
dup 173 /guilsinglright put
dup 174 /fi put
dup 175 /fl put
dup 177 /endash put
dup 178 /dagger put
dup 179 /daggerdbl put
dup 180 /periodcentered put
dup 182 /paragraph put
dup 183 /bullet put
dup 184 /quotesinglbase put
dup 185 /quotedblbase put
dup 186 /quotedblright put
dup 187 /guillemotright put
dup 188 /ellipsis put
dup 189 /perthousand put
dup 191 /questiondown put
dup 193 /grave put
dup 194 /acute put
dup 195 /circumflex put
dup 196 /tilde put
dup 197 /macron put
dup 198 /breve put
dup 199 /dotaccent put
dup 200 /dieresis put
dup 202 /ring put
dup 203 /cedilla put
dup 205 /hungarumlaut put
dup 206 /ogonek put
dup 207 /caron put
dup 208 /emdash put
dup 225 /AE put
dup 227 /ordfeminine put
dup 232 /Lslash put
dup 233 /Oslash put
dup 234 /OE put
dup 235 /ordmasculine put
dup 241 /ae put
dup 245 /dotlessi put
dup 248 /lslash put
dup 249 /oslash put
dup 250 /oe put
dup 251 /germandbls put
readonly def
currentdict end
currentfile eexec
M#1H5 J6o\&rtS(#v\0\V7Fe༙OYx5-cY9^%Y6
(efIOldN"wȰ;)UƽoFg/rvV$JS8_oc@
o9-eGJp}[Do]mN=4*{ܯq=eU^t6¤)wRfxpc@X$UČc@:p:20eַl&^
ZXs/ 1/eP6hJ/
A&;Mg*JIWCz5*E)X3l]|44R\ΙU~d 46^g1rd͂_LSg%[JedA )AdM f?i8֦1
r=Er
PP0~y9ATcs%ak.Hl
fsVY]KEЂ0'/^{>E^2u-`r^9noT+ꜮxjpA3̕Xf~utJI͇=47~>K=ڞ_`6p6^CN>3~OA73-pqNnL o\%q^|ý㭤ro,B)/TgH#R#K/p_ӿ7EO"w
Pe4.43j̼¬`Fi(!gR;.H`:֢tL$xu5*u}NkBa^}Z$H_{HnQMj'E6