CHO细胞MAR片段的克隆及其逆转录载体的构建  

Construction of CHO cells MAR sequence and construction of Retrovirus vector

作　　者：昝玉玺[1] 张俊河[1] 王天云[1] 

机构地区：[1]新乡医学院生物化学与分子生物学教研室,河南新乡453003

出　　处：《山东医药》2009年第18期18-19,共2页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(30470030);河南省自然科学基金资助项目(0511042300);河南省科技攻关项目(0624410041)

摘　　要：目的构建重组人促红素(CHO)细胞核基质结合区MAR的逆转录载体。方法以CHO细胞DNA为模板,采用PCR扩增CHO细胞的MAR序列,克隆入逆转录表达载体PLXSN-CAT中,经限制性内切酶酶切及DNA序列分析鉴定目的基因。结果PCR扩增的特异性片段长度为476 bp,以此构建的重组质粒PLXSN-CAT-MAR经BamHⅠ酶切后显示为5.9 kb和476 bp左右的两条片段,测序结果与Gen-bank中的CHO细胞MAR基因(Genbank序列号M62716)序列一致。结论成功构建了CHO细胞MAR片段PLXSN-CAT-MAR重组逆转录表达载体。Objective To construct a Retrovirus vector of CHO ceils MAR. Methods The sequence of CHO ceils MAR was amplified by polymerase chain reaction （PCR） method applied to human DNA. The fragment was inserted into Retrovirus vector PLXSN-CAT piasmid. The recombinant p].asmid was verified by double digestion and DNA sequeneing. Results The length of specific fragment applied by PCR was 500bp, and the recombinant plasmid PLXSN-CAT-MAR presented two bands ： 5.9 kb and 500bp using respective restriction enzymes BamH I. The sequence of MAR was con- firmed by blasting to Genbank. It suggested that MAR had been cloned into PLXSN-CATR vector correctly. Conclusion The recombinant Retrovirus vector PLXSN-MAR was successfully constructed.

关 键 词：核基质结合区 基因克隆 逆转录表达载体 

分 类 号：R782[医药卫生—口腔医学]