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猿猴病毒40型活缺失突变体的构建与分析

Construction and analysis of viable deletion mutants of simian virus 40.

作者信息

Shenk T E, Carbon J, Berg P

出版信息

J Virol. 1976 May;18(2):664-71. doi: 10.1128/JVI.18.2.664-671.1976.

Abstract

Viable mutants of simian virus 40 (SV40), with deletions ranging in size from 15 to 200 base pairs, have been obtained by infecting CV-1P cells with circularly permuted linear SV40 DNA. The linear DNA was produced by cleavage of closed circular DNA with DNase I in the presence of Mn2+, followed, in some cases, by mild digestion with lambda 5'-exonuclease. The SV40 map location and the size of each deletion were determined by using the S1 nuclease mapping procedure (Shenk et al., 1975) and the change in size of fragments produced by Hind II + III endonuclease cleavage. Deletions in at least three regions of the SV40 chromosome have slight or no effect on the rate or yield of viral multiplication and on vira-induced cellular transformation. These regions are located at the following coordinates on the SV40 physical map: 0.17 to 0.18; 0.54 to 0.59; and 0.68 to 0.74.

摘要

通过用环形排列的线性猴病毒40(SV40)DNA感染CV-1P细胞,已获得了SV40的存活突变体,其缺失大小范围为15至200个碱基对。线性DNA是通过在Mn2+存在下用DNase I切割闭环DNA产生的,在某些情况下,随后用λ 5'-外切核酸酶进行温和消化。通过使用S1核酸酶图谱分析程序(申克等人,1975年)以及Hind II + III内切核酸酶切割产生的片段大小变化,确定了SV40图谱位置和每个缺失的大小。SV40染色体至少三个区域的缺失对病毒增殖的速率或产量以及病毒诱导的细胞转化几乎没有影响或没有影响。这些区域在SV40物理图谱上的坐标如下:0.17至0.18;0.54至0.59;以及0.68至0.74。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ca4/515594/af07ef98d215/jvirol00221-0308-a.jpg

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