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卤虫泌壳腺特异表达蛋白SGEG1的原核表达

发表时间:2019-08-30  浏览量:147  下载量:6
全部作者: 张梦,沈帅祺,徐海晶,钱国英,马文明,王志江
作者单位: 浙江万里学院生物与环境学院;浙江医药高等专科学校制药工程学院
摘 要: 研究将卤虫泌壳腺特异表达基因1(shell gland-specifically expresses gene 1,SGEG1)的成熟肽序列克隆到带有6×His标签的原核表达载体pET-28α(+)中,构建含pET-28α(+)-SGEG1的重组质粒,转化至大肠杆菌Escherichia coli BL21(DE3)中,诱导表达重组蛋白。通过对诱导的异丙基-β-D-硫代半乳糖苷(IPTG)浓度、诱导时间和蛋白电泳检测方式的优化,确定最佳表达条件和蛋白电泳检测方法。本研究成功构建了pET-28α(+)-SGEG1重组载体,实现了分子量约为10 ku的小分子重组SGEG1蛋白的高效表达,其融合蛋白主要以包涵体形式存在,为开展SGEG1蛋白的体外制备奠定了基础。
关 键 词: 生物工程;卤虫SGEG1;原核表达;融合蛋白
Title: Prokaryotic expression of SGEG1 protein from Artemia diapause cysts
Author: ZHANG Meng, SHEN Shuaiqi, XU Haijing, Qian Guoying, MA Wenming, WANG Zhijiang
Organization: College of Biological and Environmental Sciences, Zhejiang Wanli University; College of Pharmaceutical Engineering, Zhejiang Pharmaceutical College
Abstract: In this study, the SGEG1 (shell gland-specifically expresses gene 1) sequence of deduced mature peptide was cloned into the prokaryotic expression vector pET-28α(+) with 6×His tag. A recombinant plasmid containing pET-28α(+)-SGEG1 was constructed and transformed into Escherichia coli BL21 (DE3). The fusion expression of recombinant SGEG1 (rSGEG1) protein was induced. By optimizing the induced IPTG concentration, induction time and the protein electrophoresis detection method, the optimal expression conditions and protein electrophoresis detection method were determined. The successfully constructed recombinant pET-28α(+)-SGEG1 with a molecular weight of about 10 ku was highly expressed, and the fusion protein mainly existed in the form of inclusion body, which supported a foundation for the preparation of SGEG1 protein in vitro.
Key words: bioengineering; Artemia SGEG1; prokaryotic expression; fusion protein
发表期数: 2019年8月第4期
引用格式: 张梦,沈帅祺,徐海晶,等. 卤虫泌壳腺特异表达蛋白SGEG1的原核表达[J]. 中国科技论文在线精品论文,2019,12(4):649-655.
 
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