在猴病毒40 DNA中产生小缺失的生化方法。
Biochemical procedure for production of small deletions in simian virus 40 DNA.
作者信息
Carbon J, Shenk T E, Berg P
出版信息
Proc Natl Acad Sci U S A. 1975 Apr;72(4):1392-6. doi: 10.1073/pnas.72.4.1392.
Abstract
A simple biochemical procedure for producing small deletions (15 to 50 base pairs) at virtually any location in simian virus 40 DNA has been developed. The steps involved are: cleavage of the closed-circular DNA to produce a linear structure followed by 5'-exonuclease digestion to expose a short single-stranded segment at each 3' end of the molecule. Mutants containing deletions at the site of the cleavage are obtained by infecting permissive monkey kidney cells with the exonuclease-treated DNA in the presence or absence of a helper DNA (depending upon whether or not the site of cleavage and therefore the deletion occurred in a gene required for vegetative multiplication). In this paper viable mutants with deletions at the HpaII endonuclease cleavage site (0.735 map position) and defective trans-complementable mutants with deletions at the EcoRI endonuclease cleavage site (0/1.0 map position) were isolated.
摘要
已开发出一种简单的生化程序，可在猿猴病毒40 DNA的几乎任何位置产生小缺失（15至50个碱基对）。所涉及的步骤如下：切割闭环DNA以产生线性结构，然后进行5'外切核酸酶消化，以在分子的每个3'末端暴露短的单链片段。通过在有或没有辅助DNA的情况下，用外切核酸酶处理的DNA感染允许的猴肾细胞，可获得在切割位点含有缺失的突变体（取决于切割位点以及因此缺失是否发生在营养繁殖所需的基因中）。本文分离出了在HpaII核酸内切酶切割位点（图谱位置0.735）有缺失的存活突变体和在EcoRI核酸内切酶切割位点（图谱位置0/1.0）有缺失的缺陷型反式互补突变体。
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J Mol Biol. 1974 Oct 15;89(1):179-93. doi: 10.1016/0022-2836(74)90169-7.
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Viable deletion mutants of simian virus 40: selective isolation by means of a restriction endonuclease from Hemophilus parainfluenzae.猿猴病毒40的存活缺失突变体：通过副流感嗜血杆菌的限制性内切酶进行选择性分离
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The isolation of simian virus 40 variants with specifically altered genomes.具有特定改变基因组的猿猴病毒40变体的分离。
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