Expression of recombinant classical swine fever virus E2 glycoprotein by endogenous Txnip promoter in stable transgenic CHO cells
文献类型： 外文期刊
作者： Feng, Lei 1 ; Chen, Li 1 ; Yun, Junwen 1 ; Cao, Xinglin 1 ;
作者机构： 1.Jiangsu Acad Agr Sci, Natl Res Ctr Engn & Technol Vet Biol, Inst Vet Immunol & Engn, Nanjing, Peoples R China
2.Jiangsu Coinnovat Ctr Prevent & Control Important, Yangzhou, Jiangsu, Peoples R China
3.Jiangsu Univ, Sch Pharm, Zhenjiang, Jiangsu, Peoples R China
关键词： CHO-dhfr-cells; dynamic expression; E2 protein; MTX; Txnip
期刊名称：ENGINEERING IN LIFE SCIENCES （ 影响因子：2.678； 五年影响因子：2.704 ）
ISSN： 1618-0240
年卷期： 2020 年 20 卷 8 期
页码：
收录情况： SCI
摘要： As the main immunogen that could stimulate neutralized antibody in pigs, recombinant E2 protein of CSFV was expressed in CHO-dhfr(-)cells driven by endogenous Txnip promoter from Chinese hamster. Different fragments of Txnip promoter were amplified by PCR from isolated genomic DNA of CHO cells and cloned into different expression vectors. Compared with CMV promoter, CHO-pTxnip-4-rE2 (F12) cell clone with the highest yield of rE2 protein was established by random insertion of the expression cassette driven by 860 bp sequences of Txnip promoter. In combination with treatment of 800 nM MTX for copy amplification of inserted expression cassette, the dynamic expression profile of rE2 protein was observed. Then inducible expression strategy of balance between viable cell density and product yield was conducted by mixed addition of 0.1 mM NADH and 0.1 mM ATP in culture medium at day 3 of batch-wise culture. It could be concluded that Txnip promoter would be a promising alternative promoter for recombinant antigen protein expression in transgenic cells.
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