Recombination of CBH Ⅱ Gene with Non-antibiotic Selected Vector and Expression and Activity Detection in Lactobacillus

ZHAO Ying1,2,SUN Zhe1,LIU Yan3,HU Hou-ying4,CHEN Da-fang5,LOU Yu-jie2,ZHANG Hong-fu1(1.State Key Lab of Animal Nutrition,Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193,China;2.College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China;3.College of Animal Science and Technology,Inner Mongolia Agricultural University,Huhhot 010018,China;4.Institute of Animal Science and Veterinarian of Jinan,Jinan 250002,China;5.Guizhou Province Livestock and Poultry Seedstock Breeding,Guiyang 550018,China)  

Recombinant plasmids pMD18-T-CBHⅡ and pW425t,prokaryotic expression shuttle vector between E.coli and Lactobacillus,were digested with SmaI and SphI enzymes respectively.The purified CBHⅡ gene was subcloned into the expression vector pW425t.Thus,the recombinant pW425t-CBHⅡ was constructed,and then was transformed into the competence thyA gene-mutant Lactobacillus DOMLaS107.Treated lysates of bacterium were loaded directly on SDS-PAGE,and approximately 49.6 kD protein was observed,and recombinant Lactobacillus can produced clear hydrolysis halos on the Congo-Red-CMC plate.

【Fund】： 国家自然科学基金项目(30500364);; 中国博士后基金项目(20060390897)
【CateGory Index】： Q784

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