Different matrix attachment regions flanking a transgene effectively enhance gene expression in stably transfected Chinese hamster ovary cells.

Fang Wang, Tian-Yun Wang, Yuan-Yuan Tang, Jun-He Zhang, Xian-Jun Yang

Department of Biochemistry and Molecular Biology, Xinxiang Medical University, Henan, People's Republic of China.

Gene 2012 May 25

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Numerous matrix attachment regions (MARs) have been used to improve transgene expression in genetic engineering, but an efficient and stable expression vector is lacking. In the present study, a vector named pCCF containing chloramphenicol acetyltransferase (CAT) reporter gene cassettes was constructed. The cassettes were flanked by a β-interferon MAR at the 5' upstream of the reporter gene cassettes, and a β-globin MAR at the 3' site. After transfecting pCCF into Chinese hamster ovary cells, the expression level of the CAT gene with a MAR was effectively increased to about 4.5-fold higher than that transfected with pCAM (containing two β-globin MARs flanking the expression cassette), and to 46.4-fold higher than that transfected with the control plasmid pCAG (without MARs). Quantitative reverse transcription polymerase chain reaction and the 2(-ΔΔCt) method were used to analyze the CAT gene relative copy numbers. The expression levels were found to be not directly proportional to the gene copy numbers when MAR elements from different sources were used. However, the presence of MARs improved the transgene copy numbers. Copyright © 2012 Elsevier B.V. All rights reserved.

Citation

Fang Wang, Tian-Yun Wang, Yuan-Yuan Tang, Jun-He Zhang, Xian-Jun Yang. Different matrix attachment regions flanking a transgene effectively enhance gene expression in stably transfected Chinese hamster ovary cells. Gene. 2012 May 25;500(1):59-62