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从C3H/HeN小鼠DNA中克隆与鼠白血病病毒相关的DNA序列并进行特征分析。

Molecular cloning and characterization of murine leukemia virus-related DNA sequences from C3H/HeN mouse DNA.

作者信息

Roblin R, Young J M, Mural R J, Bell T E, Ihle J N

出版信息

J Virol. 1982 Jul;43(1):113-26. doi: 10.1128/JVI.43.1.113-126.1982.

Abstract

Ten murine leukemia virus (MuLV)-related DNA sequences were isolated from C3H/HeN mouse genomic DNA by cloning of EcoRI fragments in a Charon 4A vector. Detailed restriction endonuclease maps of four of the clones were developed by using AKR MuLV [32P]cDNA as a probe. C3H clone 14-9 contains approximately 7 kilobase pairs of MuLV-related DNA, one copy of an MuLV long terminal repeat-like sequence, and a region of flanking mouse DNA. C3H clones 34.2 and 36.1 contain approximately 2 kilobase pairs of MuLV-related DNA, one copy of a MuLV LTR-like sequence, and differing lengths of flanking mouse DNA sequences. C3H clone 8.13 was found to contain an insert of 5.7 kilobase pairs of MuLV-related DNA with two long terminal repeat-like regions and sequences which are partially homologous to AKv-1. Comparison fo the restriction endonuclease cleavage maps of these C3H clones with maps recently developed for ecotropic and xenotropic MuLV DNAs indicates that C3H clone 14-9 corresponds to the 5'-terminal portion of a genomic DNA sequence related to xenotropic MuLVs, whereas C3H clones 34.2 and 36.1 correspond to the 3' terminal portions of genomic DNA sequences related to xenotropic MuLVs. Clone 8.13 represents a deleted, xenotropic MuLV-related provirus. C3H clones 14-9, 34.2, 36.1, and 8.13 provide defined DNA sequence probes with which to characterize the organization and expression of endogenous MuLV-related DNA sequences in the mouse genome.

摘要

通过将EcoRI片段克隆到Charon 4A载体中,从C3H/HeN小鼠基因组DNA中分离出10个鼠白血病病毒(MuLV)相关的DNA序列。使用AKR MuLV [32P]cDNA作为探针,绘制了其中4个克隆的详细限制性内切酶图谱。C3H克隆14 - 9包含约7千碱基对的MuLV相关DNA、一个MuLV长末端重复样序列的拷贝以及一段侧翼小鼠DNA区域。C3H克隆34.2和36.1包含约2千碱基对的MuLV相关DNA、一个MuLV LTR样序列的拷贝以及不同长度的侧翼小鼠DNA序列。发现C3H克隆8.13包含一个5.7千碱基对的MuLV相关DNA插入片段,带有两个长末端重复样区域以及与AKv - 1部分同源的序列。将这些C3H克隆的限制性内切酶切割图谱与最近为嗜亲性和异嗜性MuLV DNA绘制的图谱进行比较表明,C3H克隆14 - 9对应于与异嗜性MuLV相关的基因组DNA序列的5'末端部分,而C3H克隆34.2和36.1对应于与异嗜性MuLV相关的基因组DNA序列的3'末端部分。克隆8.13代表一个缺失的、异嗜性MuLV相关的前病毒。C3H克隆14 - 9、34.2、36.1和8.13提供了明确的DNA序列探针,可以用来表征小鼠基因组中内源性MuLV相关DNA序列的组织和表达情况。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6e/256102/a692e6e8b5a4/jvirol00154-0129-a.jpg

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