CXCR4短发夹shRNA表达质粒的构建

Construction and sequence analyzing of the recombinant plasmid affecting gene CXCR4 translation by RNA jnterfering

目的构建趋化因子受体CXCR4shRNA质粒重组体,并进行序列分析,为下一步探索肿瘤基因治疗的新途径打好基础。方法设计有小发夹结构的两条DNA序列。经退火成互补双链,再克隆至Psilencer2.1U6neo质粒载体中构建重组体,转化DH5а菌株,提取质粒行酶切鉴定后,进行测序分析。结果成功构建了CXCR4hRNA质粒重组体。结论CXCR4shRNA质粒重组体的成功构建为研究CXCR4靶向RNA干扰抗肿瘤的作用打下基础。

Objective To construct the recombinant plasmid carrying shRNA to CXCR4 and analyze the nucleic acid sequence for further searching new gene therapy method of tumor.Methods Two DNA sequences containing small hairpin structure were designed and synthesized.The complement form was obtained by annealing and inserted into vector Psilencer2.1-U6 neo,and the recombinant plasmid was transformed into DH5a strain.Finally the plasmid identified by restriction enzyme Was used for sequence analysis.Results The recombinant psilencer2.1-U6 neo carrying shRNA to CXCR4 had been constructed and the aim sequence had been obtained.Conclusion The construction of the recombinant plasmid carrying shRNA to CXCR4 lays the basis for the study of its inhibitive effect on tumor.