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水痘-带状疱疹病毒DNA的分子克隆与物理图谱分析

Molecular cloning and physical mapping of varicella-zoster virus DNA.

作者信息

Straus S E, Owens J, Ruyechan W T, Takiff H E, Casey T A, Vande Woude G F, Hay J

出版信息

Proc Natl Acad Sci U S A. 1982 Feb;79(4):993-7. doi: 10.1073/pnas.79.4.993.

Abstract

Varicella-zoster virus (VZV) DNA was cleaved with restriction endonuclease EcoRI, and most of the resulting fragments were successfully cloned in the phage vector lambda gtWES . lambda B. Double digestions of cloned fragments with EcoRI and BamHI and hybridizations to blot-transferred BamHI digests of VZV DNA were used to construct a physical map of the genome. The molecular termini of the DNA were identified by restriction enzyme analysis after exonuclease III digestion. The data indicate that VZV DNA exists in two isomeric forms that differ by inversion of one short terminal genome segment. Electron microscopic studies revealed that the short genome segment consists of a terminal revealed that the short genome segment consists of a terminal sequence of about 3.4 X 10(6) daltons that is separated from an internal inverted repeat of itself by a 5.8 X 10(60)-dalton unique DNA segment.

摘要

水痘带状疱疹病毒(VZV)DNA用限制性内切酶EcoRI切割,产生的大多数片段成功克隆到噬菌体载体λgtWES.λB中。用EcoRI和BamHI对克隆片段进行双酶切,并与VZV DNA的印迹转移BamHI酶切产物杂交,以构建基因组物理图谱。用核酸外切酶III消化后,通过限制性酶切分析鉴定DNA的分子末端。数据表明,VZV DNA以两种异构体形式存在,它们因一个短的末端基因组片段的倒位而不同。电子显微镜研究表明,短基因组片段由一个约3.4×10⁶道尔顿的末端序列组成,该序列通过一个5.8×10⁶⁰道尔顿的独特DNA片段与其自身的内部反向重复序列分开。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6350/345885/2d929c1f104a/pnas00443-0061-a.jpg

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