乳酸乳球菌食品级双筛选标记的分泌型表达载体的构建

doi:10.12160/j.issn.1672-5190.2020.02.014

畜牧与饲料科学 ›› 2020, Vol. 41 ›› Issue (2): 66-77.doi: 10.12160/j.issn.1672-5190.2020.02.014

乳酸乳球菌食品级双筛选标记的分泌型表达载体的构建

苏少锋, 萨初拉, 王潇, 吴青海, 其布日, 高娃, 赵俊利, 刘红葵, 王蕴华, 呼和

内蒙古自治区农牧业科学院,内蒙古呼和浩特 010031

收稿日期:2020-01-16 出版日期:2020-02-29 发布日期:2020-04-10
通讯作者: 呼和（1962—）,男,研究员,博士,博士生导师,主要研究方向为微生物生物技术。
作者简介:苏少锋（1984—）,男,助理研究员,博士,主要研究方向为微生物生物技术。
基金资助:
国家公益性（农业）科研专项“北方作物秸秆饲用化利用技术研究与示范”（201503134）; 内蒙古农牧业科学院青年创新基金项目（2019QNJJM04）; 内蒙古农牧业科技创新基金项目（2011CXJJM01-3）

Construction of Lactococcus lactis Food-grade Double-screening-labeled Secretory Expression Vector

SU Shao-feng, Sachula, WANG Xiao, WU Qing-hai, Qiburi, Gaowa, ZHAO Jun-li, LIU Hong-kui, WANG Yun-hua, Huhe

Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences,Hohhot 010031,China

Received:2020-01-16 Online:2020-02-29 Published:2020-04-10

摘要/Abstract

摘要： 纤维素酶在畜牧业、饲料工业、生物技术和生物能源利用方面均有广泛应用。纤维素酶基因工程作为研究的重点,可以提高酶产量和稳定性,具有广阔的应用前景。通过利用基因工程技术改造pMG36e载体,成功构建出具有双筛选标记的外分泌表达纤维素酶的食品级重组质粒,完成载体构建;然后将该质粒转化到粪肠球菌19855株并实现基因表达,经测定,分泌的纤维素酶活力为0.92 U/mL。该研究结果为后续食品级工程菌的进一步研制奠定了基础。

关键词: 纤维素酶, 载体构建, 基因表达, 食品级工程菌

Abstract: Cellulase is widely used in animal husbandry industry, feed industry, biotechnology and bioenergy utilization. Cellulase gene engineering, as one of the hot topics in biological fermentation researches, can improve enzyme yield and stability and has broad application prospects. The pMG36e vector was transformed by using genetic engineering technology, and a food-grade recombinant plasmid with double-selected markers and external secretion function of cellulase was successfully constructed. The plasmid was then transformed into Enterococcus faecalis 19855 strain and gene expression was determined. The secreted cellulase activity was 0.92 U/mL. Our results lay a foundation for the further development of food-grade engineering bacteria.

Key words: cellulose, vector construction, gene expression, food-grade engineering bacteria

中图分类号:

Q786

苏少锋, 萨初拉, 王潇, 吴青海, 其布日, 高娃, 赵俊利, 刘红葵, 王蕴华, 呼和. 乳酸乳球菌食品级双筛选标记的分泌型表达载体的构建[J]. 畜牧与饲料科学, 2020, 41(2): 66-77.

SU Shao-feng, Sachula, WANG Xiao, WU Qing-hai, Qiburi, Gaowa, ZHAO Jun-li, LIU Hong-kui, WANG Yun-hua, Huhe. Construction of Lactococcus lactis Food-grade Double-screening-labeled Secretory Expression Vector[J]. Animal Husbandry and Feed Science, 2020, 41(2): 66-77.

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乳酸乳球菌食品级双筛选标记的分泌型表达载体的构建

Construction of Lactococcus lactis Food-grade Double-screening-labeled Secretory Expression Vector

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