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目的以真核表达载体单纯疱疹病毒I型(Herpes simplex virus type 1, HSV-1)质粒型载体(pHSV)作为表达载体,构建大鼠MeCP2基因反义表达载体。方法用PCR方法扩增出大鼠MeCP2 cDNA部分片段,将其反向插入表达载体pHSV,并以PCR、酶切电泳以及DNA测序等方法对重组质粒进行鉴定。结果经过PCR、酶切电泳以及DNA测序,鉴定出连接方向和插入序列正确的反义表达载体。结论成功构建了大鼠MeCP2基因HSV-1型反义表达载体,为进一步研究该基因的功能打下了基础。
OBJECTIVE: To construct a rat MeCP2 antisense expression vector using the eukaryotic expression vector Herpes simplex virus type 1 (HSV-1) plasmid vector (pHSV) as an expression vector. Methods The partial fragment of MeCP2 cDNA was amplified by PCR and inserted into expression vector pHSV. The recombinant plasmids were identified by PCR, restriction enzyme digestion and DNA sequencing. Results After PCR, restriction enzyme digestion and DNA sequencing, the antisense expression vector with the correct orientation and insertion sequence was identified. Conclusion The HSV-1 antisense expression vector of rat MeCP2 gene was successfully constructed, which laid the foundation for further study on the function of this gene.