中文摘要：

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。

英文摘要：

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.