【摘要】 目的:探讨放射诱导肿瘤凋亡进而治疗肿瘤的新方法,构建含Egr-1启动子和Bax基因的表达载体。方法:以Egr-1启动子和Bax为靶基因,以pIRES-EGFP质粒为载体,根据Egr-1基因的DNA序列和Bax基因的c-DNA序列设计引物,分别从bab/c雌鼠的肝细胞提取含Egr-1启动子序列的DNA,从乳腺癌细胞(MCF-7)中提取含Bax-α基因序列目的基因,克隆到空载体pIRES-EGFP中,并转化至DH5α感受态细胞中,提取质粒,进行PCR鉴定和测序确认重组载体构建成功。结果:经PCR鉴定筛出的重组体的测序结果与目的基因序列相同,重组体构建成功。结论:利用克隆技术可成功构建放射调控Bax基因靶向表达的pEgr-Bax载体。更多还原

【Abstract】 Objective: To investigate a new oncotherapy from radiation-inducible apoptosis by designing and constructing expression vectors of recombinant targeting gene Egr-1 promoter and Bax. Methods: Two pairs of oligonucleotides were synthesized. Egr-1 promoter was amplified from genomic DNA of BALB/c mouse. Bax-α was amplified from MCF-7(breast-cancer cell). Egr-1 promoter and Bax-α were inserted in plasmid pIRES-EGFP. DH5α strains were transformed, plasmid was extracted, and the recombinant sequences were identified. Results: The results of recombinant sequences were the same as aim sequences by sequence identification. The recombinant was constructed successfully. Conclusion: pEgr-Bax can be constructed successfully by clonal technique.更多还原