人类心脏发育候选基因KLHL31RNAi逆转录病毒载体系统
的构建与鉴定
周军媚;叶湘漓;王跃群
【期刊名称】《湖南师范大学学报（医学版）》 【年(卷),期】2011(008)001
【摘要】Objective To construct a pSUPER RNAi vector that can inhibit human heart developmental candidate gene KLHL31 gene expression. Methods A pair of 60nt oligonucleotides coding for short hairpin RNA and targeting human heart developmental candidate gene KLHL31 were chemically synthesized and annealed and inserted into pSUPER plasmids digested with Bgl II and Xho I to construct the recombinant pSUPER RNAi plasmid (pSUPER -KLHL31). Recombinant pSUPER-KLHL31 plasmid was identified by enzyme digestion and sequencing analysis. The packaging cell H9c2 was transfected with the recombinant plasmid. Western blotting was used to determine the expression of the KLHL31 protein. Results The result of enzyme digestion and sequencing analysis demonstrated that 60 nt oligonucleotides had been inserted successfully into the vector. Green fluorescent was detected in virus- producing cells. Western blotting showed that pSUPER- KLHL31 can inhibit the expression of KLHL31 protein. Conclusion The pSUPER RNAi vector targeting human heart developmental candidate gene KLHL31 are successfully constructed and it can establish a foundation for our
人类心脏发育候选基因KLHL31RNAi逆转录病毒载体系统的构建与鉴定
