兽疫链球菌ATCC39920可控诱导表达系统的构建及其应用

生物技术通报 ›› 2016, Vol. 32 ›› Issue (12): 130-136.

兽疫链球菌ATCC39920可控诱导表达系统的构建及其应用

郑小娥¹,王震²,刘浩¹

1. 天津科技大学生物工程学院教育部工业发酵微生物重点实验室，天津 300457；
2. 中国科学院天津工业生物技术研究所系统微生物工程重点实验室天津 300308

收稿日期:2016-03-03 出版日期:2016-12-25 发布日期:2016-12-07
作者简介:郑小娥，女，研究方向：微生物遗传育种；E-mail：390857354@qq.com
基金资助:
天津市国际科技合作项目（13RCGFSY19400），天津市高等学校科技发展基金项目（20130602）

Construction of Controlled Inducible Expression System in Streptococcus zooepidemicus ATCC39920 and Its Application

ZHENG Xiao-e¹,WANG Zhen²,LIU Hao¹

1. Key Laboratory of Industrial Fermentation Microbiology，College of Biotechnology，Tianjin University of Science and Technology，Tianjin 300457；
2. Key Laboratory of Systems Microbial Biotechnology，Tianjin Institute of Industrial Biotechnology，Chinese Academy of Sciences，Tianjin 300308

Received:2016-03-03 Published:2016-12-25 Online:2016-12-07

摘要/Abstract

摘要： 旨在构建一种在兽疫链球菌（Streptococcus equi subsp. zooepidemicus）中可用的基因诱导表达系统，并探究其在两步发酵中的应用。分别构建蔗糖、乳糖、木糖和乳酸链球菌素（nisin）诱导表达载体，以编码透明质酸合成酶的hasA基因作为报告基因，以hasA基因功能缺失导致荚膜缺陷的?hasA作为宿主菌，观察诱导后荚膜生成情况验证系统的可行性。结果显示，木糖和nisin系统不能诱导hasA表达，乳糖系统诱导表达效率低，蔗糖系统可严谨且高效诱导基因表达。以?hasA/pLH201菌株实施两步发酵，葡萄糖为唯一糖源时仅用于菌体生长，添加蔗糖诱导产生透明质酸，终产量达到3.4 g/L。成功构建了一种以蔗糖为诱导物的可控-诱导基因表达系统，可用于兽疫链球菌两步发酵。

关键词: 兽疫链球菌, 可控诱导表达系统, 可控两步发酵, 透明质酸

Abstract: This work aimed at establishing an inducible gene expression system in Streptococcus zooepidemicus，and investigating the application of it in controllable two-stage fermentation. Induced expression vectors by sucrose，lactose，xylose and nisin were constructed，then using hasA of hyaluronic acid（HA）synthase as reporter gene，and ?hasA with capsule-deletion resulted from the function deletion of gene hasA as the host bacterium，the feasibility of these systems was verified by observing the generation of capsule. The results showed that：The expression of hasA was not induced in the xylose and nisin vector，at low-efficiency in lactose vector，and efficient and rigorous in sucrose vector. A controllable two-stage fermentation mode was conducted using ΔhasA/pLH201 as strain and glucose only as sole sugar source for the growth of bacteria，and added sucrose induced the generation of HA，and HA production reached at 3.4 g/L. In conclusion，this study successfully constructed a sucrose-induced gene expression system that can be used for two-stage fermentation of S. zooepidemicus.

Key words: Streptococcus zooepidemicus, controlled inducible expression system, controllable two-stage fermentation, hyaluronic acid

中图分类号:

10.13560/j.cnki.biotech.bull.1985.2016.12.021

郑小娥,王震,刘浩. 兽疫链球菌ATCC39920可控诱导表达系统的构建及其应用[J]. 生物技术通报, 2016, 32(12): 130-136.

ZHENG Xiao-e,WANG Zhen,LIU Hao. Construction of Controlled Inducible Expression System in Streptococcus zooepidemicus ATCC39920 and Its Application[J]. Biotechnology Bulletin, 2016, 32(12): 130-136.

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