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共表达ETEC K99、K88菌毛蛋白重组干酪乳杆菌的构建
引用本文:温丽娟,王桂华,侯喜林,余丽芸,王萌.共表达ETEC K99、K88菌毛蛋白重组干酪乳杆菌的构建[J].黑龙江八一农垦大学学报,2011,23(3):34-39.
作者姓名:温丽娟  王桂华  侯喜林  余丽芸  王萌
作者单位:1. 黑龙江八一农垦大学生命科学技术学院,大庆,163319
2. 黑龙江八一农垦大学动物科技学院
3. 黑龙江八一农垦大学生命科学技术学院,大庆,163319;黑龙江八一农垦大学动物科技学院
基金项目:国家人事部留学回国人员科技项目,农垦总局攻关项目
摘    要:将PCR扩增的K99和K88-LTB基因片段串联插入到干酪乳杆菌细胞表面表达载体pLA中,构建了重组表达载体pLA-K99-K88-LTB,将其电转化干酪乳杆菌CICC 6 105,获得了表达融合蛋白pLA-K99-K88-LTB的重组干酪乳杆菌表达系统.重组干酪乳杆菌在MRS培养基中表达后,经SDS-PAGE检测,约...

关 键 词:产肠毒素性大肠杆菌  K88菌毛蛋白  K99菌毛蛋白  干酪乳杆菌

The Surface Display of ETEC K99,K88 Protein in Lactobacillus casei
Wen Lijuan,Wang Guihua,Hou Xilin,Yu Liyun,Wang Meng.The Surface Display of ETEC K99,K88 Protein in Lactobacillus casei[J].Journal of Heilongjiang August First Land Reclamation University,2011,23(3):34-39.
Authors:Wen Lijuan  Wang Guihua  Hou Xilin  Yu Liyun  Wang Meng
Institution:1(1.College of Life Science and Technology,Heilongjiang Bayi Agricultural University,Daqing 163319;2.College of Animal Science and Technology,Heilongjiang Bayi Agricultural University)
Abstract:Lactobacillus casei CICC 6105 was selected as antigen delivery vehicle for the expression of enterotoxigenic Escherichia coli(ETEC) protective antigen K99 and K88 protein.The gene fragments encoding the K99 fimbrial protein and the K88-LTB fusion protein were correspondingly cloned into the Lactobacillus casei surface expression vector pLA and then the recombinant plasmid(pLA-K99-K88-LTB) was electrotransformed into Lactobacillus casei CICC6 105,resulting in recombinant strain pLA-K99-K88-LTB/L.casei.The recombinant strains were induced to express interest protein,which was detected by Western blotting,immunofluorescence microscopy and flow cytometry.The results showed that a molecular weight of about 90 kDa was detected by SDS-PAGE and the result of Western-blot indicated that the expressed protein possessed the antigenic specificity which could be recognized by mouse anti-K99 or K88 serum.The fluorescence microscopy and flow cytometric analysis showed the fusion protein was expressed on the cell surface of L.casei using PgsA,a membrane-anchored protein display motif.
Keywords:enterotoxigenic escherichia coli  K99 fimbrial protein  K88 fimbrial protein  lactobacillus casei
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