ATX短发夹shRNA表达载体的构建和测序

Construction and sequence analysis of the recombinant plasmid affecting gene ATX translation by RNA interference

目的构建ATX基因重组表达载体.并进行测序鉴定,为下一步探索肿瘤基因治疗的途径打下基础。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Psilencer2.1-U6 neo质粒载体中构建重组表达载体,转化DH5a菌株,提取质粒行酶切鉴定后,进行序列鉴定。结果成功构建了ATX shRNA重组表达载体。结论ATX shRNA质粒表达载体的成功构建为研究ATX靶向RNA干扰抗肿瘤的作用打下基础。

Objective To construct the recombinant plasmid carrying shRNA to ATX and analyze the nucleic acid sequence for further searching new gene therapy method of tumor. Methods Two DNA sequences containing short 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 cartying shRNA to ATX had been constructed and the aim sequence had been obtained. Conclusion The construction of the recombinant plasmid carrying shRNA to ATX lays the basis for the study of its inhibitive effect on tumor.