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猿猴病毒40序列在转化细胞DNA中的排列。

The arrangement of simian virus 40 sequences in the DNA of transformed cells.

作者信息

Botchan M, Topp W, Sambrook J

出版信息

Cell. 1976 Oct;9(2):269-87. doi: 10.1016/0092-8674(76)90118-5.

Abstract

High molecular weight DNA, isolated from eleven cloned lines of rat cells independently transformed by SV40, 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 SV40 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 DNAs of seven of the cell lines contained viral sequences in fragments of many different sizes. The remaining four 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.

摘要

从经SV40独立转化的11个大鼠细胞克隆系中分离出高分子量DNA,用各种限制性内切核酸酶进行切割。通过琼脂糖凝胶电泳对DNA进行分级分离,原位变性,按照Southern(1975年)所述直接转移到硝酸纤维素膜上,并与体外标记至高比活度的SV40 DNA杂交。通过放射自显影确定病毒序列在转化细胞DNA片段中的位置。7个细胞系的DNA在许多不同大小的片段中含有病毒序列。其余4个细胞系每个在不同的染色体位置含有单个病毒DNA插入片段。病毒序列与细胞序列之间的连接点在病毒基因组的不同位置定位。

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