整合到牛肿瘤细胞基因组中的牛白血病病毒DNA的分子克隆

Molecular cloning of bovine leukemia virus DNA integrated into the bovine tumor cell genome.

作者信息

Sagata N, Ogawa Y, Kawamura J, Onuma M, Izawa H, Ikawa Y

出版信息

Gene. 1983 Dec;26(1):1-10. doi: 10.1016/0378-1119(83)90030-6.

Abstract

The bovine leukemia virus (BLV) DNA harbored in the bovine tumor cell genome was cloned in lambda Charon 4A phage. Using either representative or 3' half-enriched BLV cDNA as a blot hybridization probe, clone lambda BLV-1 was shown to carry 9 kb of the BLV genome, flanked by cellular sequences at both ends. Restriction mapping with twelve endonucleases and hybridization of the DNA fragments to BLV cDNA representing a 3'-end portion of the viral genome revealed the presence and precise location of two long terminal repeats (LTRs) and virus-cell junctions. Thus, lambda BLV-1 appears to contain the complete BLV genome and flanking tumor cellular sequences. The restriction map of the cloned BLV proviral DNA closely resembles that previously reported for unintegrated linear proviral DNA, but differs significantly from that of the integrated provirus of another BLV isolate, the difference occurring preferentially in the putative gag and pol genes.

摘要

牛肿瘤细胞基因组中携带的牛白血病病毒（BLV）DNA被克隆到λ噬菌体Charon 4A中。使用代表性的或3'半富集的BLV cDNA作为印迹杂交探针，克隆的λBLV-1被证明携带9kb的BLV基因组，两端均侧翼有细胞序列。用12种内切核酸酶进行限制性图谱分析，并将DNA片段与代表病毒基因组3'端部分的BLV cDNA杂交，揭示了两个长末端重复序列（LTR）和病毒-细胞连接点的存在及其精确位置。因此，λBLV-1似乎包含完整的BLV基因组和侧翼的肿瘤细胞序列。克隆的BLV前病毒DNA的限制性图谱与先前报道的未整合线性前病毒DNA的图谱非常相似，但与另一种BLV分离株的整合前病毒的图谱有显著差异，这种差异优先发生在假定的gag和pol基因中。