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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 E1A gene was amplified by using the plasmid pE1A as a template, and then the E1A cDNA fragment was inserted between the SalI and NotI of the MCSB of the eukaryotic expression vector pIRES-Rb94 which had been constructed in the previous stage to obtain the eukaryotic double gene expression plasmid plRES-Rb94-E1A . After transfected A549 cells with lung adenocarcinoma, the expression of Rb94 and E1A in A549 cells was detected by RT-PCR and Western blot, which laid a good experimental foundation for the study of combined gene therapy for lung cancer and provided the basis for clinical combination gene therapy for lung cancer Scientific basis.