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猿猴病毒40感染细胞中的脱氧核糖核酸复制。II. 猿猴病毒40假病毒颗粒的检测与特性分析。

Deoxyribonucleic acid replication in simian virus 40-infected cells. II. Detection and characterization of simian virus 40 pseudovirions.

作者信息

Levine A J, Teresky A K

出版信息

J Virol. 1970 Apr;5(4):451-7. doi: 10.1128/JVI.5.4.451-457.1970.

Abstract

Purified simian virus 40 (SV40) virions, grown in primary African green monkey kidney cells labeled prior to infection with (3)H-thymidine, contain a variable quantity of (3)H-labeled deoxyribonucleic acid (DNA). This DNA is resistant to deoxyribonuclease, sediments at 250S, and is enclosed in a particle that can be precipitated with SV40-specific antiserum. DNA-DNA hybridization experiments demonstrate that this (3)H-labeled component in purified SV40 virions is cellular DNA. When this (3)H-labeled DNA is released from purified virus with sodium dodecyl sulfate, it has an average sedimentation constant of 14S. Sedimentation through neutral and alkaline sucrose gradients shows that this 14S DNA is composed of a collection of different sizes of DNA molecules that sediment between 11 and 15S. As a result of this size heterogeneity, SV40 virions containing cellular DNA (pseudovirions) have a variable DNA to capsid protein ratio and exhibit a spectrum of buoyant densities in a CsCl equilibrium gradient. Pseudovirions are enriched, relative to true virions, on the lighter density side of infectious SV40 virus banded to equilibrium in a CsCl gradient. Little or no cellular DNA was found in purified SV40 virus preparations grown in BSC-1 or CV-1 cells.

摘要

在感染前用³H - 胸腺嘧啶标记的原代非洲绿猴肾细胞中培养的纯化猿猴病毒40(SV40)病毒粒子,含有数量可变的³H标记的脱氧核糖核酸(DNA)。这种DNA对脱氧核糖核酸酶有抗性,在250S处沉降,并且被包裹在一个能用SV40特异性抗血清沉淀的颗粒中。DNA - DNA杂交实验表明,纯化的SV40病毒粒子中的这种³H标记成分是细胞DNA。当用十二烷基硫酸钠从纯化的病毒中释放出这种³H标记的DNA时,其平均沉降常数为14S。通过中性和碱性蔗糖梯度沉降显示,这种14S DNA由一系列不同大小的DNA分子组成,这些分子在11至15S之间沉降。由于这种大小异质性,含有细胞DNA的SV40病毒粒子(假病毒粒子)具有可变的DNA与衣壳蛋白比率,并且在CsCl平衡梯度中呈现出一系列浮力密度。相对于真正的病毒粒子,假病毒粒子在CsCl梯度中平衡离心的感染性SV40病毒带的较轻密度一侧富集。在BSC - 1或CV - 1细胞中培养的纯化SV40病毒制剂中几乎没有发现细胞DNA。

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