CD147 shRNA质粒表达载体的构建及鉴定

目的构建表达细胞外基质金属蛋白酶诱导因子(CD147,EMMPRIN)基因序列特异性的短发夹样RNA(short hairpin RNA,shRNA)载体。方法根据CD147基因序列及shRNA设计原则,化学合成两段编码短发夹RNA寡核苷酸序列,将其定向克隆到pGenesil-1.1质粒表达载体中,重组构建RNAi质粒,并对重组质粒进行酶切分析。结果限制性内切酶Eco31I和SacI酶切显示设计合成的shRNA编码序列被成功插入pGenesil-1.1质粒载体中,酶切结果证实插入片段与设计序列完全一致,成功构建针对基因CD147的shRNA质粒表达载体。结论成功构建人CD147基因重组载体,为研究CD147蛋白的生物学效应和骨肉瘤的治疗提供实验依据。

Construction and identification of CD147 shRNA expression vector

Objective To construct the expressing vector of short hairpin RNA （ shRNA） sections targeting human CD147 gene. Methods Two pairs of specific CD147 shRNA oligoes were designed and synthesized according to CD147 cDNA sequence and the principle of shRNA designing. The complement form was cloned into plasmid vector pGenesil-1.1, identified by digestion and tested for the sequence. Results The recombinant pGenesil-1.1 plasmid was cloned and the CD147 cDNA sequence was obtained. The digestion of plasmid vectors identified that DNA template primer was successfully inserted into the plasmid and the sequence was in conformity with the designed result. Conclusion Successful construction of human CD147 shRNA expressing vectors provides a favorable foundation for further study on the function of CD147.