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[目的]构建VEGF121腺相关病毒载体,为VEGF121的体外表达及临床研究提供实验基础。[方法]RT-PCR获取人VEGF121基因克隆入pAAV-MCS载体中,构建pAAV-VEGF121表达载体,利用双酶切及DNA测序鉴定pAAV-VEGF121载体的构建。[结果]RT-PCR扩增出一条444bp的特异性泳带,双酶切结果表明成功构建pAAV-VEGF121载体,DNA测序结果与VEGF121 mRNA一致。[结论]成功构建pAAV-VEGF121载体,为VEGF基因的进一步作用机制研究打下实验基础。
[Objective] To construct VEGF121 adeno-associated virus vector and provide the experimental basis for the in vitro expression and clinical study of VEGF121. [Methods] The human VEGF121 gene was cloned into pAAV-MCS vector by RT-PCR to construct pAAV-VEGF121 expression vector. The construction of pAAV-VEGF121 vector was confirmed by double enzyme digestion and DNA sequencing. [Result] A 444 bp specific PCR band was amplified by RT-PCR. The results of double enzyme digestion showed that pAAV-VEGF121 vector was successfully constructed and the result of DNA sequencing was consistent with that of VEGF121 mRNA. [Conclusion] The successful construction of pAAV-VEGF121 vector lay the experimental foundation for the further study of the mechanism of VEGF gene.