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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (30): 196-200.doi: 10.11924/j.issn.1000-6850.2012-0407

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

重组植物甜蛋白马宾灵的食品级乳酸乳球菌诱导表达系统的构建

顾文亮 夏启玉 姚晶 胡新文 符少萍 郭建春   

  • 收稿日期:2012-02-14 修回日期:2012-05-21 出版日期:2012-10-25 发布日期:2012-10-25
  • 基金资助:

    马宾灵基因(Mabinlin)体外甜味表达及其结构发生规律研究

Construction of food-grade inducible expression system for recombinant plant sweet protein MabinlinⅡ in Lactococcus lactis

  • Received:2012-02-14 Revised:2012-05-21 Online:2012-10-25 Published:2012-10-25

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摘要：

为了将马宾灵(MabinlinⅡ)直接应用于功能性食品中，通过对MabinlinⅡ基因进行有目的的剪切重组，将重组MabinlinⅡ基因插入到食品级表达载体中，经电击转化和乳糖筛选获得重组的食品级乳酸乳球菌，构建了重组植物甜蛋白马宾灵(MBL-ABH)的食品级乳酸乳球菌诱导表达系统。结果表明，该食品级诱导表达系统经乳酸链球菌素(Nisin)诱导后所表达的目的蛋白的含量较低，Western-blot检测表明MBL-ABH成功的在食品级乳酸乳球菌中表达。本研究扩展了植物甜蛋白马宾灵在食品方面的应用，有望开发出能够避免添加食糖类物质的功能性食品。

关键词: 聚合酶链式反应, 聚合酶链式反应

Abstract:

In this study, in order to directly use MabinlinⅡ in functional food, the MabinlinⅡ gene was reconstructed and then the recombinant MabinlinⅡ gene was inserted into the food-grade expression vector. After electroporation and lactose utilization screening, the food-grade inducible expression system for recombinant plant sweet protein MabinlinⅡ in L. lactis was constructed. The expression of the induced recombinant MabinlinⅡ was in a low level. The western blotting results demonstrated that the recombinant MabinlinⅡ protein was successfully expressed by the recombinant food-grade L. lactis when the inducible expression system was induced by Nisin. This research showed the feasibility of constructing a food-grade L. lactis dairy product strategy by removing the need for addition of sugar and contributing to the value of functional food.