抗端粒酶M1RNA核酶真核表达载体构建及肝癌细胞转染的研究
M1RNA核酶,,],M1RNA核酶；端粒，末端转移酶；肝癌细胞；转染,1材料与方法,2结果,3讨论,[参考文献]
[摘要] 目的 构建抗人端粒酶RNA(hTR)的M1RNA核酶的真核表达载体，并筛选稳定转染核酶的肝癌细胞株。方法 设计并应用PCR技术合成hTR模板的M1RNA核酶，应用pEGFPC1载体构建M1RNA核酶的真核表达质粒，应用脂质体LipofectAMINEAM2000转染肝癌细胞系HepG2，应用G418筛选稳定表达核酶的细胞株。结果 测序证实hTR的M1RNA核酶基因被正确克隆入真核表达载体pEGFPC1，应用脂质体成功转染肝癌细胞系HepG2。结论 成功构建了hTR的M1RNA核酶，并筛选出了稳定表达核酶的肝癌细胞株。[关键词] M1RNA核酶；端粒，末端转移酶；肝癌细胞；转染
CONSTRUCTION OF EUKARYOTIC VECTOR OF M1RNA RIBOZYME AGAINST TELOMERASE RNA AND TRANSFECTION TO HEPATOCELLULAR CARCINOMA CELL LINES
JIANG YINGJUN, CHENG GUANG, KONG XINJUAN, et al
(Department of Emergency General Surgery, The Affiliated Hospital of Qingdao University Medical College, Qingdao 266003, China)
[ABSTRACT]ObjectiveTo construct a eukaryotic expression vector of M1RNA ribozyme against telomerase RNA and transfect to hepatocellular carcinoma cell lines. MethodsM1RNA ribozyme targeting the RNA template of telomerase was designed. The eukaryotic plasmids of M1RNA ribozyme were constructed using pEGFPC1 vector. The plasmids were introduced into hepatocellular carcinoma cells (HepG2) by LipofectAMINEAM2000. ResultsThe eukaryotic expression vector of M1RNA ribozyme against telomerase RNA was constructed successfully by sequencing analysis. The hepatocellular carcinoma cell lines HepG2 was transfected successfully by M1RNA ribozyme that targeted the RNA component of telomerase. ConclusionThe stable transfected cells with M1RNA ribozyme is constructed successfully. ......
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