【摘要】 目的获得携带针对人CD59基因p SUPER retro RNAi逆转录病毒载体的卵巢癌细胞株A2780。方法设计能转录产生靶向CD59小发夹RNA(sh RNA)转染逆转录病毒载体p SUPER retro neo+gfp质粒,构建p SUPER-si CD59重组质粒,将重组质粒导入卵巢癌细胞株A2780内。鉴定转染后细胞株能形成稳定的扩增。结果重组质粒经PCR凝胶电泳、核苷酸测序证实与设计序列一致;重组质粒转染卵巢癌细胞株A2780,可稳定表达绿色荧光蛋白。结论携带针对人CD59基因p SUPER retro RNAi逆转录病毒载体的卵巢癌细胞株A2780的建立为深入进行肿瘤细胞研究奠定了基础。更多还原

【Abstract】 Objective To construct and screen the stable cell clone of p SUPER gene retro RNAi of CD59 in A2780 by using the expression vector of si RNA. Methods 60 bp oligonucleotide sequences targeting CD59 RNA were cloned into p SUPER retro neo + gfp plasmid,and the recombinant plasmid was transfected into ovarian cancer cell line A2780. The stable cell clones can be formed after the transfection. Results The recombinant vector was identified by PCR and restriction enzyme digestion. The recombinant vector was transfected into ovarian cancer cell line A2780,which can be stably expressed in green fluorescent protein. Conclusions p SUPER retro RNAi of CD59 gene in A2780 was established as a basis for further study on human ovarian cancer.更多还原