[1]陈帅印),张荣光),范清堂),等.乳酸菌中幽门螺杆菌ureB基因的食品级表达与优化*[J].郑州大学学报(医学版),2011,(03):396.
 CHEN Shuaiyin),ZHANG Rongguang),FAN Qingtang),et al. Foodgrade expression of helicobacter pylori ureB gene in Lactococcus lactis and optimization for its expression conditions[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2011,(03):396.
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乳酸菌中幽门螺杆菌ureB基因的食品级表达与优化*
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2011年03期
页码:
396
栏目:
论著
出版日期:
2011-05-15

文章信息/Info

Title:
 Foodgrade expression of helicobacter pylori ureB gene in Lactococcus lactis and optimization for its expression conditions
作者:
 陈帅印1)张荣光2)范清堂1) 段广才1)#
1)郑州大学公共卫生学院流行病学教研室;河南省分子医学重点实验室 郑州 450001
2)郑州大学基础医学院寄生虫学教研室 郑州 450001
Author(s):
CHEN Shuaiyin1)ZHANG Rongguang2)FAN Qingtang1)DUAN Guangcai1)
1)Department of Epidemiology,College of Public Health,Zhengzhou University; Henan Key Laboratory of Molecular Medicine,Zhengzhou 450001
2)Department of Parasitology,College of Basic Medical Sciences,Zhengzhou University,Zhengzhou 450001
关键词:
幽门螺杆菌ureB基因乳酸乳球菌食品级表达载体条件优化
分类号:
R183
文献标志码:
A
摘要:
目的:以乳酸乳球菌(Lactococcus lactis, L. lactis)为宿主菌构建幽门螺杆菌(Helicobacter pylori,H. pylori)尿素酶B(UreB)的食品级表达系统。方法:从重组质粒pMD19TureB上酶切得到ureB基因片段,与含有筛选标记lacF 基因的L. lactis表达载体pNZ8149经双酶切后定向连接,应用PCR、酶切和测序的方法鉴定重组子。重组菌用乳联菌肽诱导表达,用正交表实验进行表达条件的优化,通过SDSPAGE和Western blot对重组蛋白进行鉴定。结果:重组菌NZ3900/pNZ8149ureB经鉴定与预期相符。优化结果显示在菌体培养到D600 nm为0.40左右时加入终浓度为25 μg/L的乳联菌肽诱导5 h产量最高,通过Western blot能检测到UreB的表达。结论:构建了ureB基因食品级表达系统,并得到了表达的最优条件。

参考文献/References:

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备注/Memo

备注/Memo:
 *中国博士后科学基金资助项目20070410252
#通讯作者,男,1958年8月生,博士,教授,研究方向:分子流行病学,E-mail: gcduan@public2.zz.ha.cn
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