MAR提高稳定转化的CHO细胞转基因表达

基础医学与临床 ›› 2008, Vol. 28 ›› Issue (11): 1125-1128.

MAR提高稳定转化的CHO细胞转基因表达

王天云韩忠敏杨献军董忠生王清义

新乡医学院细胞生物化学与分子生物学教研室郑州铁路职业技术学院医学院郑州铁路职业技术学院医学院

收稿日期:2007-11-20 修回日期:2007-12-21 出版日期:2008-11-25 发布日期:2008-11-25
通讯作者: 王天云

MAR Increased Transgene Expression in Stable Transfected CHO Cells

Tian-yun WANG, Zhong-min HAN, Xian-jun YANG, Zhong-sheng DONG, Qing-yi WANG

Dep. of Biochemistry and Molecular Biology, Xinxiang Medical University Medical College of Zhengzhou Railway Vocational College

Received:2007-11-20 Revised:2007-12-21 Online:2008-11-25 Published:2008-11-25
Contact: Tian-yun WANG,

摘要/Abstract

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（Chloramphenicol acetyltransferase,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能提高转基因CAT的表达水平5.5倍，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。

关键词: 核基质结合区, 转基因, 基因工程, CHO细胞

Abstract: Objective To study the effect of human β-globin matrix attachment region (MAR) on transgene expression in stably transfected CHO cells. Methods The eapression vector was constructed , which contained the β-globin MAR in both sides of Chloramphenicol acetyltransferase (CAT) reporter gene expression cassette in cis, then transfected into CHO cells and screened under G418 selection. The CAT reporter gene expression was analyzed through ELISA method. Results It has showed that theβ-globin MAR could enhance 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 could increase transgene expression in stably transfected CHO cells.

Key words: matrix attachment region, transgene, gene engineering, CHO Cells

王天云韩忠敏杨献军董忠生王清义. MAR提高稳定转化的CHO细胞转基因表达[J]. 基础医学与临床, 2008, 28(11): 1125-1128.

Tian-yun WANG; Zhong-min HAN; Xian-jun YANG; Zhong-sheng DONG; Qing-yi WANG. MAR Increased Transgene Expression in Stable Transfected CHO Cells[J]. Basic & Clinical Medicine, 2008, 28(11): 1125-1128.

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