ARRANGEMENT OF SIMIAN-VIRUS 40 SEQUENCES IN DNA OF TRANSFORMED-CELLS
ARRANGEMENT OF SIMIAN-VIRUS 40 SEQUENCES IN DNA OF TRANSFORMED-CELLS
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DOI：
10.1016/0092-8674(76)90118-5
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发表时间：
1976-01-01
期刊：
影响因子：
64.5
通讯作者：
SAMBROOK, J
中科院分区：
文献类型：
--
作者：
BOTCHAN, M;TOPP, W;SAMBROOK, J
从11个经SV-40独立转化的大鼠细胞克隆系中分离出高分子量DNA，用不同的限制性内切酶进行切割。DNA通过琼脂糖凝胶电泳分离，原位变性，按照Southern（1975）的描述直接转移到硝化纤维素片上，并与体外标记为高比活性的SV-40 DNA杂交。病毒序列在转化细胞DNA片段中的位置由放射自显影确定。其中7个细胞系的DNA含有不同大小的病毒序列片段。剩下的4个细胞系，每个细胞系在不同的染色体位置都包含一个病毒DNA的插入。病毒和细胞序列之间的连接点位于病毒基因组的不同位置。
High molecular weight DNA, isolated from eleven cloned lines of rat cells independently transformed by SV-40, was cleaved with various restriction endonucleases. The DNA was fractionated by electrophoresis through agarose gels, denatured in situ, transferred directly to sheets of nitrocellulose as described by Southern (1975) and hybridized to SV-40 DNA labeled in vitro to high specific activity. The location of viral sequences among the fragments of transformed cell DNA was determined by autoradiography. The DNA of 7 of the cell lines contained viral sequences in fragments of many different sizes. The remaining 4 cell lines each contain a single insertion of viral DNA at a different chromosomal location. The junctions between viral and cellular sequences map at different places on the viral genome.