莫洛尼氏小鼠肉瘤病毒未整合DNA在噬菌体λ中的分子克隆
Molecular cloning of unintegrated Moloney mouse sarcoma virus DNA in bacteriophage lambda.
作者信息
Verma I M, Lai M H, Bosselman R A, McKennett M A, Fan H, Berns A
出版信息
Proc Natl Acad Sci U S A. 1980 Apr;77(4):1773-7. doi: 10.1073/pnas.77.4.1773.
The covalently closed circular forms of unintegrated viral DNA obtained from cells infected with Moloney mouse sarcoma virus was cloned in bacteriophage lambda. The viral DNA was cleaved with restriction endonuclease HindIII and inserted in the unique HindIII site of lambda Charon 21A DNA. Recombinant clones containing virus-reactive DNA sequences were analyzed by restriction endonuclease mapping, R-loop formation, and infectivity assays. Two of eight genome-length recombinant clones characterized contained the large terminal repeat. Only the recombinant clones containing the large terminal repeat were able to induce focus formation in uninfected mouse fibroblasts.
从感染莫洛尼氏小鼠肉瘤病毒的细胞中获得的未整合病毒DNA的共价闭合环状形式被克隆到噬菌体λ中。病毒DNA用限制性内切酶HindIII切割，并插入到λ噬菌体21A DNA的独特HindIII位点。通过限制性内切酶图谱分析、R环形成和感染性测定来分析含有病毒反应性DNA序列的重组克隆。在八个已鉴定的基因组长度重组克隆中，有两个含有大的末端重复序列。只有含有大末端重复序列的重组克隆能够在未感染的小鼠成纤维细胞中诱导灶形成。
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