沉默人p38MAPK基因表达的RNA干扰的逆转录病毒载体的构建  

Construction of a RNA interfering retrovirus vector for making silence of p38MAPK gene expression

作　　者：朱美华[1] 温红艳[1] 唐普润[1] 梁敏[1] 刘启才[2] 

机构地区：[1]广州医学院第二附属医院,广东广州510260 [2]广州医学院实验医学研究中心,广东广州510182

出　　处：《现代中西医结合杂志》2008年第30期4667-4668,4693,共3页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：广东省医学科研基金项目(A2006306)

摘　　要：目的构建沉默人p38MAPK基因表达的RNA干扰的逆转录病毒表达载体。方法查找人p38MAPK cDNA序列,利用软件设计并合成能够形成互补双链并能转录出靶向人p38MAPK基因的短发夹状RNA的两条寡核苷酸序列,退火后连接至线性化载体pSIREN上,获得重组质粒pSIREN-p38MAPK,酶切和测序进行鉴定。结果双酶切重组质粒获得特定的酶切图谱,证实所合成核酸片段的正确重组;测序结果显示重组核酸片段序列与设计的序列完全相同。结论成功构建了沉默人p38MAPK基因的重组质粒pSIREN-p38MAPK。Objective It is to approach the method of constructing a RNA interfering retrovirus vector for making silence the expression of p38MAPK gene.Methods Search for the sequence of human p38MAPK cDNA,design and synthesize with software two short hairpin-shape oligonucleotide sequence that can form a complementary double-strain and transcript target human p38MARK gene,after annealing,connecting to the linear vector of pSIREN,harvesting a recombinant plasmid pSIREN-p38MAPK,identification through enzyme digestion and sequencing was made.Results Specific restriction enzyme map was obtained from double enzyme digestion,verified correctness of the recombination of synthesized nucleate fragment;sequencing showed identity between the recombinant nucleate fragment and the designed sequence.Conclusion The recombinant plasmid pSIREN-p38MAPK that can make silence the human p38MAPK gene is successfully constructed.

关 键 词：RNA干扰 P38MAPK 逆转录病毒表达载体 

分 类 号：R-33[医药卫生]