绿色荧光蛋白穿梭载体的构建及其在嗜酸乳杆菌中的应用

doi:10.16431/j.cnki.1671-7236.2015.10.006

Abstract

Abstract: In order to further study the mechanism of Lactobacillus acidophilus,GFP labeling shuttle expression vector pSET4s-P1-GFP-P2 was constructed by inserting green fluorescence protein (GFP),upstream and downstream homology arm of Lactobacillus acidophilus based on temperature shuttle expression vector pSET4s.Recombinant Lactobacillus acidophilus expressing green fluorescence gene was constructed by transforming pSET4s-P1-GFP-P2 into Lactobacillus acidophilus using electrotransformation.We could obtain the recombinant Lactobacillus acidophilus by resistance screening and temperature screening,and named it as ΔMG6243(GFP) which could expressed GFP gene stably by fluorescence microscopy observation and Western blotting analysis.We could analyse the genetic stability and biological characteristics with fluorescence microscopy observation and plate count.Green fluorescent of recombinant Lactobacillus acidophilus could be observed in the blue light,and also be observed after 10 generations.There were significant differences neither growth characteristics nor pH and salt tolerance compared with the wild mushrooms.The results showed that GFP labeling recombinant Lactobacillus acidophilus was constructed successfully.Exogenous gene could be non-resistant and integrative to express in the Lactobacillus acidophilus with the vector.It established the foundation for construction of recombinant Lactobacillus acidophilus.At the same time,the role of the GFP as marker laid the foundation for distribution,adhesion of probiotic in animal gastrointestinal tract and the mechanism of Lactobacillus acidophilus.

Key words: Lactobacillus acidophilus; homologous recombination; green fluorescent protein

CLC Number:

ZENG Juan-juan, WANG Lei, ZHAO Di, LV Yang, WANG Lei, YI Dan, CHEN Hong-bo, DING Bin-ying, HOU Yong-qing, WU Tao. Construction of the Shuttle Vector Expressing Green Fluorescent Protein and its Application in Lactobacillus acidophilus[J]. , 2015, 42(10): 2551-2559.

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Construction of the Shuttle Vector Expressing Green Fluorescent Protein and its Application in Lactobacillus acidophilus

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