Gilmer T M, Parsons J T
J Virol. 1979 Dec;32(3):762-9. doi: 10.1128/JVI.32.3.762-769.1979.
The cellular sites of integration of avian sarcoma virus (ASV) have been examined in clones of duck embryo cells infected with the Bratislava 77 strain of ASV using restriction endonuclease digestion, agarose gel electrophoresis, Southern blotting, and hybridization with labeled ASV complementary DNA probes. DNA prepared from 11 clones of duck embryo cells infected with the Bratislava 77 strain of ASV was digested with the restriction enzymes HpaI, which cleaves once within the viral genome, and Hind III, which cleaves twice within the viral genome, and the virus-cell DNA juncture fragments were resolved by agarose gel electrophoresis. Analysis of the virus-cell junctures present in individual ASV-infected duck embryo clones revealed that all clones contain at least one copy of nondefective proviral DNA with some clones containing as many as 5 to 6 copies of proviral DNA. A comparison of the virus-cell juncture fragments present in different ASV-infected clones showed that each clone contains a unique set of virus-cell junctures. These data suggest that ASV DNA can integrate at multiple sites within the duck embryo cell genome and that these sites appear to be different as defined by digestion with the restriction enzymes HpaI and HindIII.
利用限制性内切酶消化、琼脂糖凝胶电泳、Southern印迹法以及与标记的禽肉瘤病毒（ASV）互补DNA探针杂交等方法，对感染了布拉迪斯拉发77株ASV的鸭胚细胞克隆中ASV的细胞整合位点进行了检测。从感染了布拉迪斯拉发77株ASV的11个鸭胚细胞克隆中提取的DNA，用限制性内切酶HpaI（在病毒基因组内切割一次）和Hind III（在病毒基因组内切割两次）进行消化，病毒-细胞DNA连接片段通过琼脂糖凝胶电泳进行分离。对单个感染ASV的鸭胚克隆中存在的病毒-细胞连接进行分析发现，所有克隆都至少含有一个非缺陷型前病毒DNA拷贝，有些克隆含有多达5至6个前病毒DNA拷贝。对不同感染ASV的克隆中存在的病毒-细胞连接片段进行比较表明，每个克隆都含有一组独特的病毒-细胞连接。这些数据表明，ASV DNA可以整合到鸭胚细胞基因组内的多个位点，并且根据用限制性内切酶HpaI和HindIII消化的结果，这些位点似乎是不同的。
