真核双基因表达载体plRES-Rb94-E1A的构建与鉴定  

Construction and Identification of the Eukaryotic Co-expression Plasmid pIRES-Rb94-E1A

作　　者：刘芳 李清宝[2] 李萌 栾力[3] 王建波[3] 程玉峰[3] 

机构地区：[1]山东医学高等专科学校影像系,山东济南250002 [2]山东大学附属省立医院心脏外科,山东济南250021 [3]山东大学齐鲁医院放疗科,山东济南250012

出　　处：《医学与哲学（B）》2011年第6期59-61,共3页Medicine & Philosophy(B)

摘　　要：以pE1A质粒为模板,扩增E1A基因,然后将E1A cDNA片段插入前期已经构建好的真核表达载体pIRES-Rb94的MCSB的SalI与NotI之间,获得真核双基因表达质粒plRES-Rb94-E1A,转染肺腺癌A549细胞后,采用RT-PCR和Western blot方法检测Rb94和E1A在A549细胞中的表达,为研究联合基因治疗肺癌奠定了良好的实验基础,且为临床联合基因治疗肺癌提供科学依据。The cDNA encoding E1A was obtained by RT--PCR from plasmid E1A, then inserted into the MCSB of the plasmid pIRES--Rb94 between SalI and NotL Plasmid pIRES--Rb94--E1A was constructed and A549 cells were trans- formed by plasmid pIRES--Rb94--E1A. RT--PCR and Western blot were used to identify the co--expression of Rb94 and E1A in A549 cells. The successful construction of plasmid pIRES--Rb94--E1A can establish an experimental basis and provide scientific foundation for clinical cancer treatment.

关 键 词：基因 表达载体 A549细胞 

分 类 号：R734.2[医药卫生—肿瘤]