MAR提高稳定转化的CHO细胞转基因表达  

MAR increased transgene expression in stable transfected CHO cells

作　　者：王天云[1] 韩忠敏[2] 杨献军[1] 董忠生[2] 王清义[2] 

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 [2]郑州铁路职业技术学院医学院,河南郑州450052

出　　处：《基础医学与临床》2008年第11期1125-1128,共4页Basic and Clinical Medicine

基　　金：国家自然科学基金(30470030);河南省科技厅自然科学基金(511042300);河南省科技厅科技攻关(0624410041)

摘　　要：目的研究MAR在CHO细胞中对转基因的表达调控作用,为建立稳定高效表达的CHO表达系统奠定基础。方法将人β-珠蛋白MAR片段顺式克隆到氯霉素乙酰转移酶(CAT)报告基因表达盒两侧,构建MAR介导的哺乳动物细胞表达载体,转染CHO细胞,G418筛选稳定表达细胞株,ELISA分析CAT报告基因的表达水平。结果β-珠蛋白MAR能提高5.5倍转基因CAT的表达水平,并可降低不同细胞转化株之间的转基因表达差异。结论MAR能提高转基因在稳定转染的CHO细胞的表达。Objective To study the effect of humanβ-globin matrix attachment region （MAR） on transgene expression in stably transfected CHO cells. Methods Expression vector was constructed, which contained the β-globin MAR in both sides of Chloramphenicol acetyltransferase （CAT） reporter gene expression cassette in c/s, then trans- letted into CHO cells. The CAT reporter gene expression was analyzed by ELISA method. Results The β-globin MAR enhanced the CAT gene expression 5.5-fold in stably transformed CHO cells, while the transgene expression variation among individuals of transformants was decreased. Conclusion MAR increase transgene expression in stably transfected CHO cells

关 键 词：核基质结合区 转基因 基因工程 CHO细胞 

分 类 号：Q782[生物学—分子生物学]