中国动脉硬化杂志2012，Vol.20Issue(5)：397-401,5.
靶向人类心脏发育候选基因hole的siRNA真核表达载体的构建和鉴定
Construction and Identification of Eukaryotic siRNA Expression Vector Targeting Human Heart Developmental Candidate Gene hole Gene
摘要
Abstract
Aim To construct a pSUPER RNAi system that targets human heart developmental candidate gene hole and a stable virus-producing cell line. Methods A pair of 60nt oligonucleotides coding for short hairpin RNA and targeting human heart developmental candidate gene hole 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-hole). Recombinant pSUPER-hole plasmid was identified by enzyme digestion and sequencing analysis. The packaging cell H9c2 was transfected with the recombinant plasmid. The tnRNA expression level of hole was detected by RT-PCR. Results The result of enzyme digestion and sequencing analysis demonstrated that 60 nt had been inserted successfully into the vector. Green fluorescent was detected in virus-producing cells, RT-PCR showed pSUPER-hole can inhibit the hole gene expression of H9c2. Conclusion The pSUPER RNAi vector targeting human heart developmental candidate gene hole are successfully constructed.关键词
心脏发育/hole/pSUPER/siRNA/H9c2细胞Key words
Heart Development/hole/pSUPER/siRNA/H9c2分类
生物科学引用本文复制引用
周军媚,姚峰,谢华平,叶湘漓,王跃群..靶向人类心脏发育候选基因hole的siRNA真核表达载体的构建和鉴定[J].中国动脉硬化杂志,2012,20(5):397-401,5.基金项目
国家自然科学基金项目(81000035、31071999和30871340) （81000035、31071999和30871340）
