猪A组轮状病毒pW425et-Vp4基因工程乳酸菌的构建及表达  

Construction and expression of recombinant pW425et-Vp4 genetic engineering Lactobacillus of porcine rotavirus A

作　　者：叶丽萍[1] 王春玲 王春凤[1] 

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118 [2]吉林省永吉县畜牧局黄榆乡畜牧站,吉林永吉132223

出　　处：《中国兽医杂志》2012年第1期3-6,共4页Chinese Journal of Veterinary Medicine

基　　金：国家863计划(2006AA10A205,2007AA10Z322);国家自然科学基金(30671573,30870116)

摘　　要：以嗜酸乳杆菌为受体菌株,将pW425et-Vp4重组质粒电转化入嗜酸乳杆菌,构建猪源A组轮状病毒Vp4基因工程乳酸菌,对影响电转化效率的主要因素进行研究,并对构建的乳酸菌进行表达分析。结果表明,在MRS培养基中加入1%甘氨酸+0.3mol/L蔗糖,电场强度为11kV/cm,脉冲时间为3.8ms,电击后细胞复苏3～4h等条件时,得到较高的转化效率,最高可达3.8×104 CFU/μg DNA。pW425et-Vp4乳酸菌阳性克隆经Western-blot分析表明,其蛋白具有与猪轮状病毒多克隆抗体的反应原性。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.

关 键 词：猪源A组轮状病毒Vp4基因 乳酸菌 电转化 表达 

分 类 号：S852.659.4[农业科学—基础兽医学]