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《Chinese Journal of Veterinary Medicine》 2012-01
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Construction and expression of recombinant pW425et-Vp4 genetic engineering Lactobacillus of porcine rotavirus A

YE Li-ping1,WANG Chun-ling2,WANG Chun-feng1 (1.College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China; 2.HuangYu Livestock Farm Station of Animal Husbandry Bureau,Yongji 132223,China)  
In order to obtain Vp4 genetic engineering Lactobacillus of rotavirus group A,pW425et-Vp4 recombinant plasmid of Porcine rotavirus was transfected into Lactobacillus acidophilus by electroporation.The electroporation efficiency was determined and the reactionogenicity of recombinant Lactobacillus was analyzed.Results showed that the optimal values for concentration of glycine and of sucrose,the electrical field strength,the pulse time and the length of revival are 1%(W/V),0.3 mol/L,11 kV/cm,3.8 ms and 3~4 h,respectively,the best electroporation efficiency is 3.8×104 CFU/μg DNA.In addition,the protein of pW425et-Vp4 recombinant Lactobacillus could be recognized by the antibody against Rotavirus A as examined by Western blotting,indicating that the protein has epitopes that can be recognized by polyclonal antibody against porcine rotavirus.
【Fund】: 国家863计划(2006AA10A205 2007AA10Z322);; 国家自然科学基金(30671573 30870116)
【CateGory Index】: S858.28
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