牛Inhibin pET32a-matpeptide重组蛋白质的诱导表达及鉴定

doi:10.11924/j.issn.1000-6850.casb16060051

中国农学通报 ›› 2017, Vol. 33 ›› Issue (14): 121-124.doi: 10.11924/j.issn.1000-6850.casb16060051

所属专题：生物技术；畜牧兽医

牛Inhibin pET32a-matpeptide重组蛋白质的诱导表达及鉴定

吴迪,穆秋玲,齐晓龙,常迪,李贺娟,郭勇,倪和民

北京农学院动物科学技术学院,吉林省白城市畜牧总站,北京农学院动物科学技术学院,北京农学院动物科学技术学院,北京农学院动物科学技术学院,北京农学院动物科学技术学院,北京农学院动物科学技术学院

收稿日期:2016-06-11 修回日期:2016-08-24 接受日期:2016-08-25 出版日期:2017-05-25 发布日期:2017-05-25
通讯作者: 倪和民
基金资助:
2013 年度北京市教委北京市属高等学校创新团队建设与教师职业发展计划项目“体细胞转基因克隆肉牛新品系培育与利用” (IDHT20130515)；奶牛产业体系北京市创新团队。

Induced Expression and Identification of Recombinant Protein of Bovine Inhibin pET32a-matpeptide

Received:2016-06-11 Revised:2016-08-24 Accepted:2016-08-25 Online:2017-05-25 Published:2017-05-25

摘要/Abstract

摘要： 为表达纯化出免疫原性良好的牛Inhibin pET32a-matpeptide重组蛋白质,本试验通过对菌液IPTG诱导时间、温度以及诱导浓度的摸索确定了蛋白诱导最佳表达条件,利用重组蛋白小量表达确定目的蛋白以可溶性或包涵体蛋白存在,重组蛋白大量表达纯化出大量目的蛋白,通过Western blot鉴定目的蛋白免疫原性。结果表明：使用1MIPTG,18℃诱导表达18h可诱导表达出重组抑制素蛋白,经重组蛋白小量表达确定目的蛋白以可溶性形式存在,Western blog试验表明重组蛋白有较高的免疫原性。本试验诱导表达出可溶性、免疫原性较高的Inhibin pET32a-matpeptide重组蛋白质,为后续制备抗抑制素α亚基抗体奠定了基础。

关键词: 脂多糖, 脂多糖, Toll样受体2, Toll样受体4, 肠粘膜微血管内皮细胞

Abstract: To expressed and purified good immune original of Bovine Recombinant Inhibin pET32a-matpeptide protein, the research induced and expressed Inhibin recombinant protein by determining the induction time, temperature and IPTG induced concentration, through the restructuring of the small protein expression to determine the target protein to soluble and inclusion body protein exists, by the recombinant protein expression to purify the target protein and by Western blot to identify the protein"s immunogenicity. The results showed that the use of 1MIPTG, 18℃ to induce the expression of 18h was induced to express the recombinant inhibin protein, the recombinant protein epsilon expression for determining protein exists in a soluble form and by Western blot to identify the protein has good immunogenicity. The Inhibin pET32a-matpeptide recombinant protein was induced to express soluble high immunogenicity, which laid the foundation for the preparation of anti-Inhibin α subunit antibody.

中图分类号:

Q812

吴迪,穆秋玲,齐晓龙,常迪,李贺娟,郭勇,倪和民. 牛Inhibin pET32a-matpeptide重组蛋白质的诱导表达及鉴定[J]. 中国农学通报, 2017, 33(14): 121-124.

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牛Inhibin pET32a-matpeptide重组蛋白质的诱导表达及鉴定

Induced Expression and Identification of Recombinant Protein of Bovine Inhibin pET32a-matpeptide

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