重组绵羊FSH在CHO-K1细胞中的表达及其应用

doi:10.13560/j.cnki.biotech.bull.1985.2019-0668

生物技术通报 ›› 2019, Vol. 35 ›› Issue (12): 112-117.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0668

重组绵羊FSH在CHO-K1细胞中的表达及其应用

王兴陇^{1, 2}, 李倩^{1, 2}, 马瑞仙^{1, 2}, 李生军^{1, 2}, 李向茸¹, 冯若飞¹

1. 西北民族大学生物医学研究中心生物工程与技术国家民委重点实验室,兰州 730030;
2. 西北民族大学生命科学与工程学院,兰州 730030

收稿日期:2019-07-23 出版日期:2019-12-26 发布日期:2019-12-03
作者简介:王兴陇,男,硕士研究生,研究方向：分子病毒学;E-mail：1406181280@qq.com
基金资助:
国家自然科学基金项目（31460665）,西北民族大学中央高校基本科研业务费专项资金资助研究生项目（Yxm2019147）,国家民委中青年英才计划[批准号（2018）98],教育部“创新团队发展计划”项目（IRT_17R88）

Expression of Recombinant Ovine FSH in the CHO-K1 Cell and Its Application

WANG Xing-long^{1, 2}, LI Qian^{1, 2}, MA Rui-xian^{1, 2}, LI Sheng-jun^{1, 2}, LI Xiang-rong¹, FENG Ruo-fei¹

1. Key Laboratory of Biotechnology & Bioengineering of State Ethnic Affairs Commission,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030;
2 College of Life Science and Engineering,Northwest Minzu University,Lanzhou 730030

Received:2019-07-23 Published:2019-12-26 Online:2019-12-03

摘要/Abstract

摘要： 在CHO-K1细胞中分泌表达重组绵羊FSH及其生物活性初探。构建重组表达质粒pcDNA3.1-FSHβα,转染至全悬浮CHO-K1细胞中,经过G418加压筛选出表达量高的单克隆细胞,于生物反应器中扩大培养,并进行超滤浓缩,将浓缩后的重组绵羊FSH免疫小鼠,测定小鼠卵巢重量并分析血清中黄体生成素和孕酮含量。成功构建了重组质粒pcDNA3.1-FSHβα,并成功在全悬浮CHO-K1细胞中实现分泌表达,表达量为297.97 ng/mL,注射重组绵羊FSH的小鼠卵巢重量显著增加,黄体生成素和孕酮含量均显著上调。重组绵羊FSH蛋白在CHO-K1细胞中成功表达,并具有一定的生物活性。

关键词: 促卵泡激素, CHO-K1细胞, 生物反应器

Abstract: This work aims to explore the expression of recombinant sheep FSH（follicle stimulating hormone）in CHO-K1 cells and its biological activity. The recombinant plasmid pcDNA3.1-FSHβα was constructed and electroporated into fully suspended CHO-K1 cells. The monoclonal cells with high expression of recombinant FSH protein were screened under G418 pressure,cultured in a bioreactor and concentrated by ultrafiltration. The concentrated recombinant sheep FSH was injected into mice,the ovarian mass of mice was weighed,and the contents of luteinizing hormone and progesterone in mouse serum were detected. The recombinant plasmid pcDNA3.1-FSHβα was successfully constructed and the secretory expression was successful in CHO-K1 cells,and the expression level was 297.97 ng/mL. The ovaries of mice injected with concentrated recombinant sheep FSH protein were significantly enlarged,and the contents of luteinizing hormone and progesterone were also significantly up-regulated. In sum,the recombinant sheep FSH protein is successfully expressed in CHO-K1 cells and presents certain biological activity.

Key words: follicular stimulating hormone, CHO-K1 cells, bioreactor

王兴陇, 李倩, 马瑞仙, 李生军, 李向茸, 冯若飞. 重组绵羊FSH在CHO-K1细胞中的表达及其应用[J]. 生物技术通报, 2019, 35(12): 112-117.

WANG Xing-long, LI Qian, MA Rui-xian, LI Sheng-jun, LI Xiang-rong, FENG Ruo-fei. Expression of Recombinant Ovine FSH in the CHO-K1 Cell and Its Application[J]. Biotechnology Bulletin, 2019, 35(12): 112-117.

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重组绵羊FSH在CHO-K1细胞中的表达及其应用

Expression of Recombinant Ovine FSH in the CHO-K1 Cell and Its Application

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