【摘要】 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。更多还原

【Abstract】 Objective: To investigate the function of MeCP2 gene during brain development, an antisense rat MeCP2 gene expression vector was constructed using Herpes simplex virus type-1 (HSV-1) plasmid vector (pHSV). Methods: The rat MeCP2 cDNA fragment was amplified from pBluescript SK-Mecp2 by PCR and inserted into pHSV in an antisense orientation. Then PCR, restriction endonulease digestion and DNA sequencing were carried out to verify the antisense expression vector. Results: The expression vector with the correct sequence in antisense orientation was finally identified by PCR, restriction endonulease digestion and DNA sequencing. Conclusion: The antisense rat MeCP2 gene expression vector is successfully constructed and might shed light on further studies of its function.更多还原