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首页> 外文期刊>Journal of Virology >Molecular cloning of Snyder-Theilen feline leukemia and sarcoma viruses: comparative studies of feline sarcoma virus with its natural helper virus and with Moloney murine sarcoma virus.
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Molecular cloning of Snyder-Theilen feline leukemia and sarcoma viruses: comparative studies of feline sarcoma virus with its natural helper virus and with Moloney murine sarcoma virus.

机译：Snyder-Thilen猫素瘤和Sarcoma病毒的分子克隆：猫苜蓿病毒与天然辅助病毒的比较研究和莫尼鼠肉瘤病毒。

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Extrachromosomal DNA obtained from mink cells acutely infected with the Snyder-Theilen (ST) strain of feline sarcoma virus (feline leukemia virus) [FeSV(FeLV)] was fractionated electrophoretically, and samples enriched for FeLV and FeSV linear intermediates were digested with EcoRI and cloned in lambda phage. Hybrid phages were isolated containing either FeSV or FeLV DNA "inserts" and were characterized by restriction enzyme analysis, R-looping with purified 26 to 32S viral RNA, and heteroduplex formation. The recombinant phages (designated lambda FeSV and lambda FeLV) contain all of the genetic information represented in FeSV and FeLV RNA genomes but lack one extended terminally redundant sequence of 750 bases which appears once at each end of parental linear DNA intermediates. Restriction enzyme and heteroduplex analyses confirmed that sequences unique to FeSV (src sequences) are located at the center of the FeSV genome and are approximately 1.5 kilobase pairs in length. With respect to the 5'-3' orientation of genes in viral RNA, the order of genes in the FeSV genome is 5'-gag-src-env-c region-3'; only 0.9 kilobase pairs of gag and 0.6 kilobase pairs of env-derived FeLV sequences are represented in ST FeSV. Heteroduplex analyses between lambda FeSV or lambda FeLV DNA and Moloney murine sarcoma virus DNA (strain m1) were performed under conditions of reduced stringency to demonstrate limited regions of base pair homology. Two such regions were identified: the first occurs at the extreme 5' end of the leukemia and both sarcoma viral genomes, whereas the second corresponds to a 5' segment of leukemia virus "env" sequences conserved in both sarcoma viruses. The latter sequences are localized at the 3' end of FeSV src and at the 5' end of murine sarcoma virus src and could possibly correspond to regions of helper virus genomes that are required for retroviral transforming functions.
机译：从肠道细胞中获得的含有豆类素（SAT）猫群病毒（猫白血病病毒）（FESV（FELV））的乳链细胞的胚胎细胞，分级电离学，并用ECORI消化富含FELV和FESV线性中间体的样品在λ噬菌体中克隆。分离杂化噬菌体含有Fesv或FELV DNA“插入物”，其特征在于限制酶分析，R型与纯化的26至32s病毒RNA，以及异拷贝形成。重组噬菌体（指定的Lambda Fesv和Lambda Felv）含有Fesv和Felv RNA基因组中所示的所有遗传信息，但缺乏750个碱基的延伸终端冗余序列，其在父母的每端出现一次。限制酶和异拷基分析证实，FESV（SRC序列）独特的序列位于FESV基因组的中心并且长度为约1.5千碱基对。关于病毒RNA中的基因的5'-3'取向，FESV基因组中基因的顺序为5'-GAG-SRC-ENV-C区-3';在ST Fesv中仅在ST Fesv中仅为0.9千碱基对Gag和0.6千碱基对的env衍生的Felv序列。在减少严格性的条件下，在减少严格的条件下进行λFesv或λFelvDNA和莫尼鼠Sarcoma病毒DNA（菌株M1）之间的异络分析，以证明基对同源性的有限区域。鉴定了两种这些区域：第一次发生在白血病的极端5'末端和肉瘤病毒基因组，而第二个则对应于在Sarcoma病毒中保守的白血病病毒“Env”序列的5'段。后一种序列在FESV SRC的3'末端局部化，在鼠肉瘤病毒SRC的5'末端，并且可能对应于逆转录病毒转化功能所需的辅助病毒基因组的区域。

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