Expression of Novel Bifunctional Glutathione Synthetase in Pichia pastoris and Escherichia coli(新型双功能谷胱甘肽合成酶的真核和原核表达)

通过裂解无乳链球菌(Streptococcus agalactiae)菌体细胞得到基因组DNA。根据GenBank 上已发表的无乳链球菌(菌种编号ATCC13813)新型双功能谷胱甘肽合成酶基因(gshF)的核酸序列与2 种不同表达载体多克隆位点序列,分别设计引物F1、R1 和F2、R2,再以总DNA 为模板,经过特异性PCR 扩增出长度约为2 200 bp 的目的基因。随后分别对目的基因gshF 和2 种表达载体进行双酶切、连接等操作,得到表达载体pPIC9K-gshF 与 pET-gshF。用Sal I 线性化表达载体pPIC9K-gshF 后电转至毕赤酵母GS115 中。经MD 平板筛选...

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Bibliographic Details
Main Authors: LIJiahui(李佳慧), YANGFangfang(杨芳芳), WANGZhen(王珍), ZHANGSainan(张赛南), WANGShoufeng(王首锋)
Format: Article
Language:zho
Published: Zhejiang University Press 2019-07-01
Series:Zhejiang Daxue xuebao. Lixue ban
Subjects:
Online Access:https://doi.org/10.3785/j.issn.1008-9497.2019.04.014