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  

HIGH-LEVEL EXPRESSION OF HBSAG IN CHO CELLS USING GLUTAMINE SYNTHETASE GENE AS AN AMPLIFIABLE SELECTABLE MARKER

作　　者：刘文军[1] 杨芙蓉[1] 阮力[1] 任贵方[1] 朱既明[1] 

机构地区：[1]中国预防医学科学院病毒学研究所

出　　处：《病毒学报》1997年第2期103-109,共7页Chinese Journal of Virology

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。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.

关 键 词：谷氨酰胺合成酶 基因 表达 乙型肝炎 表面抗原 

分 类 号：Q753[生物学—分子生物学] R512.620.3[医药卫生—内科学]