【摘要】 目的:构建和鉴定大鼠卡氏肺孢子(Pneumocystis carinii,PC)胸腺嘧啶核苷酸合酶(Thymidylate synthase,TS)基因的siRNA(Short interfering RNA)表达载体。方法:人工合成针对PCTS基因的1对shRNA(Short hairpin RNA)序列并定向克隆到空载体pTZU6+1上,构建siRNA表达重组质粒pPC-TS,并采用双酶切产物凝胶电泳和基因测序法鉴定。结果:酶切电泳和基因测序鉴定得到的产物与预期的目的基因一致。结论:成功构建卡氏肺孢子胸腺嘧啶核苷酸合酶基因siRNA表达载体。更多还原

【Abstract】 Objective:To construct and identify a plasmid vector of short interfering RNA(siRNA) on pneumocystis carinii (PC) thymidylate synthase. Methods: Short hairpin RNA oligonucleotides of thymidylate synthase were chemically synthesized and inserted into plasmid vector pTZU6+1 after annealing. The recombinant plasmid,pPC-TS,transformed into E. coil. TOP10 and amplified,was digested by restriction endonucleases Hind Ⅲ and EcoR Ⅰ and identified by gel electrophoresis and DNA sequencing. Results: Gel electrophoresis and DNA sequencing showed that the recombinant plasmid containing the correct and full shRNA oligonucleotide. Conclusion:The siRNA plasmid,pPC-TS,was constructed successfully.更多还原