利用反义RNA探索外源基因丢失的分子机理

STUDIES ON THE MECHANISM OF DELETION OF EXOGENOUS GENES BY THE USE OF ANTISENSE RNA TECHNIQUE

利用反义RNA技术研究了调控PARP酶基因的表达对外源基因整合稳定性的影响。将PARP基因cDNA的部分序列反向插入到真核表达载体pSMG中，将重组质粒分别导入携带有外源基因的细胞中，地塞米松诱导反义PARP基因的表达后，进行Southern杂交检测。结果表明，外源基因仍保留在基因组中，这意味着外源基因的丢失并不是由于单一PARP酶活性降低所致。

The effects of inducible expression of poly (ADP-ribose) polymerase antisense RNA on deletion of the exogenous genes in NIH3T3 and HeLa cells were studied. A recombinant expression plasmid was prepared with the MMTV promoter upstream of the antisense-oriented partial PARP cDNA. After induction of antisense RNA expression by dexamethasone for the abundant concentration of PARP, the genonic DNA in both NIH3T3 and HeLa cells exhibited corresponding exogenous bands by Southern blot analysis. The results indicated that the deletion of the exogenous genes induced by inhibitors of PARP may be not only due to the reduction of PARP activity.