Distance effect of matrix attachment regions on transgene expression in stably transfected Chinese hamster ovary cells

The β-globin matrix attachment regions (MARs) were inserted into the 5′-site of the eukaryotic expression vector cassette and DNA fragments 350 and 750 bp in length were inserted into the site to generate expression vectors with varying distances between the expression cassette and MAR. The vectors...

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Veröffentlicht in:Biotechnology letters 2014-10, Vol.36 (10), p.1937-1943
Hauptverfasser: Zhang, Jun-He, Wang, Xiao-Yin, Wang, Tian-Yun, Wang, Fang, Dong, Wei-Hua, Wang, Li, Zhao, Chun-Peng, Chai, Shu-Jie, Yang, Rui, Li, Qin
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Sprache:eng
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Zusammenfassung:The β-globin matrix attachment regions (MARs) were inserted into the 5′-site of the eukaryotic expression vector cassette and DNA fragments 350 and 750 bp in length were inserted into the site to generate expression vectors with varying distances between the expression cassette and MAR. The vectors containing MARs increased chloramphenicol acetyltransferase (CAT) expression levels compared to the negative control vector lacking the MAR; the highest expression increase was 3.8-fold. A greater MAR-transgene distance (750 bp) correlated with a greater increase in transgene expression when compared to the control vector that lacked separation between the MAR and transgene. CAT gene copy numbers were higher in cells transformed with the vector possessing a smaller MAR-transgene distance (350 bp) than in cells belonging to the other three groups. However, MAR-induced transgene expression levels did not exhibit a direct relationship with gene copy number.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-014-1563-9