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利用谷氨酰胺合成酶基因(GS)作扩增标记在CHO细胞中高效表达乙型肝炎表面抗原基因
引用本文:刘文军,杨芙蓉,阮力,任贵方,朱既明.利用谷氨酰胺合成酶基因(GS)作扩增标记在CHO细胞中高效表达乙型肝炎表面抗原基因[J].病毒学报,1997(2).
作者姓名:刘文军  杨芙蓉  阮力  任贵方  朱既明
作者单位:中国预防医学科学院病毒学研究所
摘    要:利用谷氨酰胺合成酶基因(GS)[1]作扩增选择标记,结合CMV-IE启动子,在CHO细胞中高效表达乙型肝炎表面抗原基因。初筛克隆表达水平RPHA检测为1:64,经过谷氨酰胺合成酶基因的抑制剂MSX的两轮基因扩增,HBsAg的表达水平RPHA在1:256以上。方瓶静置培养收液,RIA检测HBsAg最高产量为9.5μg/毫升。表达水平较以前利用dhfr基因扩增选择系统所得到的高表达细胞系B43高一倍以上。利用GS基因扩增选择系统可以在哺乳动物细胞中高水平表达外源基因。

关 键 词:GS基因扩增系统,HBsAg,CHO细胞,基因基达

HIGH-LEVEL EXPRESSION OF HBSAG IN CHO CELLS USING GLUTAMINE SYNTHETASE GENE AS AN AMPLIFIABLE SELECTABLE MARKER
Liu Wenjun\ Yang Furong\ Ruan li\ Ren Guifang\ Zhu Jiming.HIGH-LEVEL EXPRESSION OF HBSAG IN CHO CELLS USING GLUTAMINE SYNTHETASE GENE AS AN AMPLIFIABLE SELECTABLE MARKER[J].Chinese Journal of Virology,1997(2).
Authors:Liu Wenjun\ Yang Furong\ Ruan li\ Ren Guifang\ Zhu Jiming
Abstract:We have reported a method for introducing glutamine synthetase gene(GS) intl CHO cells in which transfectants are selected by growth in a glutamine-free medium.Vector amplification can be subsequently selected using specific inhibitor of GS,hte methionine sulphoximine(MSX).Using this system,combining with CMV promoter,HBV S gene were expressed in CHO cells.Af ter two rounds of selection for vector amplification,the expression level of G4 cell line is above 1:256(RPHA),as much as twice of the B43 cell line,which we obtained using dhfr gene amplification.The GS amplification system has the advantage of selecting high expression cell line more easily.
Keywords:GS gene amplification system  HBsAg  CHO cell  Gene expression
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