CTGF靶向RNA干扰重组体的构建及序列分析  

Construction and sequence analysis of connective tissue growth factor targeted recombinant plasmids by RNA interference

作　　者：主余华[1] 张春清[1] 赵幼安[2] 褚衍六[2] 孙成刚[1] 

机构地区：[1]山东大学山东省立医院消化内科,山东省济南市250021 [2]山东大学齐鲁医院消化内科,山东省济南市250012

出　　处：《世界华人消化杂志》2005年第24期2880-2883,共4页World Chinese Journal of Digestology

基　　金：山东省卫生厅青年基金资助项目;No.2005-25~~

摘　　要：目的:利用pEGFP质粒载体构建介导结缔组织生长因子(CTGF)短发夹RNA(shorthairpinRNA,shRNA)表达的质粒.方法:分别设计3对有小发夹结构的两条DNA序列,经退火成互补双链,再克隆至带有U6启动子的质粒载体pEGFP中,构建重组体,转化DH5α菌株,提取质粒行酶切鉴定后,进行测序分析.结果:CTGF的sRNA片段被成功克隆到pEGFP质粒载体中,3个重组质粒shRNA编码序列与设计的片段完全一致,经酶切与测序证实构建成功.结论:成功构建了能表达CTGFshRNA的重组体,为下一步探索肝纤维化基因治疗的新途径打好基础.AIM： To construct the recombinant plasmids expressing connective tissue growth factor （CTGF） short hairpin RNA （shRNA） by pEGFP plasmid vector. METHODS： Three pairs of two DNA sequences containing small hairpin structure were designed and synthesized respectively, and then were formed into complementary chains by annealing. The obtained products were inserted into plasmid vector pEGFP containing U6 promoter. Then the recombinant plasmids were transformed into DH5α strain. Finally the plasmids that identified by restriction enzyme were used for sequence analysis. RESULTS： The CTGF shRNA expression frames were successfully inserted into the plasmid vector pEGFP, and the shRNA coding sequences of the 3 obtained recombinant plasmids were consistent with the designed fragments. The cloned recombinants were identified and confirmed by endonuclease digestion and sequence analysis. CONCLUSION： The recombinant plasmids of shRNA for CTGF are successfully constructed.

关 键 词：结缔组织生长因子 RNA干扰 短发夹RNA 质粒 

分 类 号：R346[医药卫生—基础医学]