Bcl xL和BDDFⅧ在CHO DG44细胞中共
表达及抗凋亡能力分析
Anti apoptosis competence of Bcl xL by co expression of BDDFⅧ in CHO/DG44 cells
作者：王继超(四川大学生命科学学院生物资源与生态环境教育部重点实验室)；聂艳桃(成都蓉生药业有限责任公司)；何太平(成都蓉生药业有限责任公司)；费保进(成都蓉生药业有限责任公司)；林宏辉(四川大学生命科学学院生物资源与生态环境教育部重点实验室)；雷韬(成都蓉生药业有限责任公司)
Author：Wang Ji Chao(Key Laboratory of Bio Resource and Eco environment of Ministry of Education,College of
Life Sciences,Sichuan University)；NIE Yan Tao(Chengdu Rongsheng Pharmaceuticals, Co., Ltd)；HE Tai Ping(Chengdu Rongsheng Pharmaceuticals, Co., Ltd)；FEI Bao Jin(Chengdu Rongsheng Pharmaceuticals, Co., Ltd)；LIN Hong Hui(Key Laboratory of Bio Resource and Eco environment of Ministry of Education,College of
Life Sciences,Sichuan University)；LEI Tao(Chengdu Rongsheng Pharmaceuticals, Co., Ltd)
收稿日期：2014-11-17 年卷（期）页码：2015,52(6):1411-1416
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：B区缺失型凝血因子Ⅷ（BDDFⅧ）； 抗凋亡； CHO细胞； Bcl xL； 表达Q81A0490
Key words：B domain deleted factor Ⅷ (BDDFⅧ)；Anti apoptosis；CHO cell; Bcl xL ;Expression
基金项目：重组人凝血因子VIII的研制(2014JY0103
中文摘要
在CMV启动子控制下构建含有B区缺失型FⅧ基因（BDDFⅧ）和Bcl xL基因编码序列的真核表达载体pF8B和只含B区缺失的FⅧ基因（BDDFⅧ）的真核表达载体pF8，通过双酶切和测序验证重组真核表达载体构建的正确性；经过MTX压力筛选得到稳定表达BDDFⅧ活性的CHO F8B和CHO F8细胞株.在相同可控培养参数下进行发酵培养，Bcl xL基因过表达赋予CHO F8B细胞较强的抗调亡能力和更高的BDDFⅧ表达活性，BDDFⅧ的表达活性后者是前者的5倍多.
英文摘要
A recombinant eukaryotic expression vector pF8B containing B domain deleted factor Ⅷ(BDDFⅧ) and Bcl xL genes ,respectively under the control of CMV promoters ,was constructed .Recombinant expression vector F8 without the Bcl xL gene has the same bone frame as above .Then they were transfected into CHO DG44 cells by lipotransfection .And transfectants were screened for stable expression cell lines ,which were analyzed for growth status and the expression of target protein. The results showed that the construction of expression vectors were correct by restriction analysis and sequencing. Cell lines CHO F8B containing BDDFⅧ and Bcl xL genes and CHO F8 with BDDFⅧ gene inside only were obtained through MTX selection.Under the same condition of controllable culture parameters ,over expression of Bcl xL gene in CHO F8B cells could make recombinant cells more robust against apoptosis resulting in increasing the expression level of interested protein in the fermentation and the expression of BDDFⅧ in CHO F8B cells was five times higher than CHO F8 cells.
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