PEDV ps420片段乳酸乳球菌表达及免疫中和活性检测  

Expressing PEDV ps420 by recombinant Lactococcus lactis and neutralizing detection after immunization

作　　者：汪淼[1] 葛俊伟[1] 姜艳平[1] 唐丽杰[1] 乔薪瑗[1] 李一经[1] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030

出　　处：《东北农业大学学报》2011年第12期20-24,I0011,共6页Journal of Northeast Agricultural University

基　　金：黑龙江十一五科技攻关项目(GA06B202-4)

摘　　要：将猪流行性腹泻病毒LJB/03株S基因上编码中ps420(1 495～1 916 bp)基因片段插入乳酸乳球菌pNZ8112中,构建了重组表达载体pNZ8112-ps420,将其电转化入乳酸乳球菌Lactococcus lactis NZ9000,获得了表达猪流行性腹泻病毒S基因ps420的重组乳酸乳球菌,经乳链菌肽(Nisin)在GM17培养基中诱导表达,SDS-PAGE检测表明,约有大小分别为15 ku的蛋白得到表达,与理论值相符。Westernblot结果和间接免疫荧光试验结果分析表明,表达的蛋白在菌体表面且可被兔源PEDV抗血清所识别。重组菌口服免疫接种家兔获得抗血清,进行PEDV细胞中和试验结果表明重组菌可诱发血清中和IgG产生,血清中和抗体效价达1∶105。Lactococcus lactis was selected as a bacterial carrier for the expression of Porcine epidemic diarrhea virus （PEDV） neutralizing epitope domain ps420. The gene encoding PEDV ps420 protein was cloned into the Lactococcus lactis expression vector pNZ8112, and then the constructed recombinant vector pNZ8112-ps420 was electrotransformed into Lactococcus lactis NZ9000 generating the recombinant system pNZ8112-ps420/NZ9000 expressing PEDV ps420 protein. The recombinant strain was induced by Nisin in GM17 and about 15 ku protein was detected with SDS-PAGE. The result of Western-blot indicated that the expressed protein possessed the antigenic specificity which could be recognized by rabbite anti-PEDV serum. The indirect immune. Rabbits were immunized via oral route with pNZ8112-ps420/NZ9000 to identify whether the recombinant strain have the ability to induce systemic antibody responses. The gene encoding PEDV ps420 protein could be certificated as a neutralizing antigen domain.

关 键 词：猪流行性腹泻病毒 中和表位区 乳酸乳球菌 中和抗原 

分 类 号：S828[农业科学—畜牧学]