论文部分内容阅读
目的 :将重构型caspase 6分子 (RCasp 6 )克隆入可诱导真核表达载体 pIND中 ,经蜕皮激素类似物诱导探讨其对细胞凋亡的促进作用。方法 :用PCR扩增RCasp 6基因 ,并克隆入真核表达载体pIND中。以重组体转染Hela细胞系 ,蜕皮激素类似物诱导。RCasp 6基因的表达产物采用免疫细胞化学染色检测。HE染色观察转染细胞的形态变化 ,并计数细胞绘制生存曲线。结果 :成功地构建了RCasp 6基因的可诱导真核表达载体。转染的细胞经蜕皮激素诱导后 ,细胞形态异常。转染了RCasp 6基因的细胞固缩同时很多细胞死亡。 结论 :RCasp 6基因具有促进细胞死亡的作用。
OBJECTIVE: To clone the reconstructed caspase 6 (RCasp 6) into the inducible eukaryotic expression vector pIND, and to explore the role of caspase 6 in promoting the apoptosis of the cell line induced by ecdysone analogues. Methods: RCasp 6 gene was amplified by PCR and cloned into eukaryotic expression vector pIND. The Hela cell lines were transfected with recombinant cells and induced by ecdysone analogs. The expression product of RCasp 6 gene was detected by immunocytochemical staining. The morphological changes of transfected cells were observed by HE staining and the survival curves of cells were counted. Results: The inducible eukaryotic expression vector of RCasp 6 gene was successfully constructed. Transfected cells induced by ecdysone, cell morphology abnormalities. Cells that have been transfected with the RCasp 6 gene shrink and many of the cells die. Conclusion: RCasp 6 gene can promote cell death.