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首页> 外文期刊>Journal of Virology >Characterization of proviruses cloned from mink cell focus-forming virus-infected cellular DNA.
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Characterization of proviruses cloned from mink cell focus-forming virus-infected cellular DNA.

机译：从水貂细胞聚焦性病毒感染细胞DNA中克隆的潜水术的表征。

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Two proviruses were cloned from EcoRI-digested DNA extracted from mink cells chronically infected with AKR mink cell focus-forming (MCF) 247 murine leukemia virus (MuLV), using a lambda phage host vector system. One cloned MuLV DNA fragment (designated MCF 1) contained sequences extending 6.8 kilobases from an EcoRI restriction site in the 5' long terminal repeat (LTR) to an EcoRI site located in the envelope (env) region and was indistinguishable by restriction endonuclease mapping for 5.1 kilobases (except for the EcoRI site in the LTR) from the 5' end of AKR ecotropic proviral DNA. The DNA segment extending from 5.1 to 6.8 kilobases contained several restriction sites that were not present in the AKR ecotropic provirus. A 0.5-kilobase DNA segment located at the 3' end of MCF 1 DNA contained sequences which hybridized to a xenotropic env-specific DNA probe but not to labeled ecotropic env-specific DNA. This dual character of MCF 1 proviral DNA was also confirmed by analyzing heteroduplex molecules by electron microscopy. The second cloned proviral DNA (designated MCF 2) was a 6.9-kilobase EcoRI DNA fragment which contained LTR sequences at each end and a 2.0-kilobase deletion encompassing most of the env region. The MCF 2 proviral DNA proved to be a useful reagent for detecting LTRs electron microscopically due to the presence of nonoverlapping, terminally located LTR sequences which effected its circularization with DNAs containing homologous LTR sequences. Nucleotide sequence analysis demonstrated the presence of a 104-base-pair direct repeat in the LTR of MCF 2 DNA. In contrast, only a single copy of the reiterated component of the direct repeat was present in MCF 1 DNA.
机译：使用λ噬菌体宿主载体系统，从慢性感染的水貂细胞中提取的Mink细胞中提取的生态消化DNA从生态消化的DNA中克隆了两种潜水术。一个克隆的Mulv DNA片段（指定的MCF 1）含有从5'长终端重复（LTR）中的ECORI限制性位点延伸6.8千比下的序列，到位于信封（ENV）区域中的ECORI位点，并且通过限制内切核酸酶测定难以区分5.1千杆间（LTR中的ECORI站点除外）来自AKR生态转化玻璃体DNA的5'末端。从5.1〜6.8千碱基延伸的DNA段含有几个不存在于AKR生态分子潜水艇的限制性位点。位于MCF 1 DNA的3'末端的0.5千碱基DNA段含有与异熵ENV特异性DNA探针杂交但不标有生态学env特异性DNA的序列。通过电子显微镜通过分析异单分子分子，还证实了MCF 1荧光DNA的这种双重特征。第二克隆的荧光DNA（指定MCF 2）是6.9千碱基的ECORI DNA片段，其在每个末端含有LTR序列，并且包含大多数ENV区域的2.0千碱基缺失。 MCF 2荧光DNA被证明是一种有用的试剂，用于检测LTR电子电子显微镜，由于存在非封闭的LTR序列，其与含有同源LTR序列的DNA进行循环化。核苷酸序列分析证明在MCF 2 DNA的LTR中存在104碱基的直接重复。相反，在MCF 1 DNA中仅存在直接重复的重复组分的单个拷贝。

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