
基本信息:
- 专利标题: 双顺反子表达载体及其构建方法与应用
- 专利标题(英):Bicstronic expression vector, and construction method and application thereof
- 申请号:CN201610284371.6 申请日:2016-05-03
- 公开(公告)号:CN107338267A 公开(公告)日:2017-11-10
- 发明人: 万晓春 , 李俊鑫 , 刘绿艳 , 任晓虎 , 周启明
- 申请人: 中国科学院深圳先进技术研究院 , 深圳先进技术研究院 , 深圳市疾病预防控制中心 , 深圳市南山区人民医院
- 申请人地址: 广东省深圳市南山区西丽大学城学苑大道1068号
- 专利权人: 中国科学院深圳先进技术研究院,深圳先进技术研究院,深圳市疾病预防控制中心,深圳市南山区人民医院
- 当前专利权人: 中国科学院深圳先进技术研究院,深圳先进技术研究院,深圳市疾病预防控制中心,深圳市南山区人民医院
- 当前专利权人地址: 广东省深圳市南山区西丽大学城学苑大道1068号
- 代理机构: 北京三友知识产权代理有限公司
- 代理人: 韩蕾; 姚亮
- 主分类号: C12N15/85
- IPC分类号: C12N15/85 ; C12N15/66
The invention provides a bicstronic expression vector, and a construction method and application thereof, and concretely relates to the bicstronic expression vector used for efficiently and stably transfecting a dihydrofolate reductase auxotrophic mutation CHO cell strain (CHO/dhfr-). The bicstronic expression vector is constructed in a way of inserting an S100a6 promotor, a target gene and a dhfr gene on a pIRES plasmid. According to the bicstronic expression vector, and the construction method and the application thereof provided by the invention, through using the low-temperature inducible promoter and constructing the bicstronic vector, the expression of the dhfr gene is weakened, and the target gene acquires sufficient copy number under low-concentration MTX (Methotrexate) only through two or three times of pressure selection; and meanwhile, by utilizing the property of the inducible promoter, the temperature is reduced during a production process, and the yield of recombinant proteins is increased maximally.