【摘要】 构建能表达 Lac Z基因和 c-Ha-T2 4 ras基因的两种真核表达重组质粒 A1 3.4和 B1 2 .7(两基因相对位置不同 ) ,并将其转染 NIH3T3细胞和 He La细胞 ,经 G4 1 8筛选 ,分别建立了四种稳定转化细胞系 A1 3.4 -NIH3T3、B1 2 .7-NIH3T3、A1 3.4 -He La及 B1 2 .7-He La.用聚 ADP核糖聚合酶 (PARP)的 NAD位点抑制剂苯甲酰胺(BA)分别处理四种转化细胞后 ,检测细胞中整合的外源 Lac Z基因与 c-Ha-T2 4 ras基因的删除情况 ,结果如下 :BA诱导的基因删除可发生于 NIH3T3转化细胞系 ,但不能发生于 He La转化细胞系 ;位于同一外源表达载体上的 Lac Z基因与 c-Ha-T2 4 ras基因的删除过程是同步的 ;外源整合 DNA片段的转录强度可能直接影响其被 BA诱导删除的敏感性更多还原

【Abstract】 Two plasmids A13.4 and B12.7 were constructed in which both c Ha T24ras gene and LacZ gene were inserted in different direction. Then four transformed cell lines, A13.4 NIH3T3, B12.7 NIH3T3, A13.4 HeLa, B12.7 HeLa were obtained by transfecting A13.4 and B12.7 into NIH3T3 and HeLa cells. After treatment of benzamide (BA) which is an inhibitor of poly (ADP ribose) polymerase (PARP), β galactosidase activity assays and PCR analysis were performed to test the deletion of LacZ gene and c Ha T24ras gene. The results showed that:(1)LacZ gene was deleted from both A13.4 NIH3T3 cells and B12.7 NIH3T3 cells, but neigher from A13.4 HeLa cells nor B12.7 HeLa cells. So it is possible that there is cell line specificity in gene deletion induced by BA. (2) In A13.4 NIH3T3 cells, LacZ gene and c Ha T24ras gene were completely deleted at the same time. (3) The transcription activity of exgenous DNA fragment may have same effect on its sensitivity to the deletion induced by BA.更多还原