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口服表达纤维素酶基因的重组乳酸乳球菌可提高鹅对纤维的消化率。

Oral Administration of Recombinant Lactococcus lactis Expressing the Cellulase Gene Increases Digestibility of Fiber in Geese.

作者信息

Zhou Haizhu, Gao Yunhang, Gao Guang, Lou Yujie

机构信息

College of Animal Science and Technology, Jilin Agricultural University, Changchun, 130118, China.

出版信息

Curr Microbiol. 2015 Dec;71(6):693-8. doi: 10.1007/s00284-015-0904-9. Epub 2015 Sep 4.

Abstract

Enhancing cellulose digestibility in animals is important for improving the utilization of forage, which can decrease the amount of food used in animal production. The aim of the present study was to achieve recombinant expression of the cellulase gene in Lactococcus lactis and evaluate the effects of oral administration of the recombinant L. lactis on fiber digestibility in geese. Cellulase (Cell) and green fluorescent protein (GFP) genes were cloned into a L. lactis expression vector (pNZ8149) to construct the recombinant expression plasmid (pNZ8149-GFP-Cell). Then, the recombinant expression plasmid was transformed into L. lactis (NZ3900) competent cells by electroporation to obtain recombinant L. lactis (pNZ8149-GFP-Cell/NZ3900) in which protein expression was induced by Nisin. Expression of GFP and Cell by the recombinant L. lactis was confirmed using SDS-PAGE, fluorescence detection, and Congo red assays. A feeding experiment showed that oral administration of pNZ8149-GFP-Cell/NZ3900 significantly increased the digestibility of dietary fiber in geese fed either a maize stalk diet or a rice chaff diet. Therefore, oral administration of recombinant L. lactis cells expressing the cellulase gene increases fiber digestibility in geese, offering a way to increase the utilization of dietary fiber in geese.

摘要

提高动物对纤维素的消化率对于提高草料利用率很重要,这可以减少动物生产中所用的饲料量。本研究的目的是在乳酸乳球菌中实现纤维素酶基因的重组表达,并评估口服重组乳酸乳球菌对鹅纤维消化率的影响。将纤维素酶(Cell)基因和绿色荧光蛋白(GFP)基因克隆到乳酸乳球菌表达载体(pNZ8149)中,构建重组表达质粒(pNZ8149-GFP-Cell)。然后,通过电穿孔将重组表达质粒转化到乳酸乳球菌(NZ3900)感受态细胞中,以获得重组乳酸乳球菌(pNZ8149-GFP-Cell/NZ3900),其中蛋白质表达由乳酸链球菌素诱导。使用SDS-PAGE、荧光检测和刚果红试验确认了重组乳酸乳球菌对GFP和Cell的表达。一项饲养实验表明,口服pNZ8149-GFP-Cell/NZ3900可显著提高以玉米秸秆日粮或稻壳日粮喂养的鹅对膳食纤维的消化率。因此,口服表达纤维素酶基因的重组乳酸乳球菌细胞可提高鹅对纤维的消化率,为提高鹅对膳食纤维的利用率提供了一种方法。

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