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《Chinese Journal of Biotechnology》 2002-03
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Construction of Eukaryotic Expression Vector Using Neomycin-resistanceGene Mutant as Selectable Marker

GAO Chuan 1* ZHU Xu-Dong 2 ZHOU Xiao-Wei 2 YU Fang 2 LU Bai-Song 2 HUANG Pei-Tang 2 1(Institute of Pharmaceutical Chemistry, Beijing 102205,China) 2(Institute of Biotechnology, Beijing 100071,China)  
Neomycin-resistance gene is widely used as a selectable marker in eukaryotic expression vector. It codes neomycin phosphotransferaseⅡ(NPTⅡ) which confers resistance to various aminoglycoside antibiotic such as G418 and kanamycine. In this work ,by site-directed mutagenesis the neo gene mutant was obtained. The expression vector pmDNA using the neo gene mutant as selectable marker has been constructed. After inserting interest luciferase gene, the expression plasmid pmDNAluc+ was stably transfected CHO-K1 cells. As a result, the expression positive ratio reaches to approximate 95% and the ratio of high expression colonies is apparently higher than the controls.
【CateGory Index】: Q782
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