中国畜牧兽医2017，Vol.45Issue(10)：2851-2857,7.DOI:10.16431/j.cnki.1671-7236.2017.10.004
山羊痘病毒毒力因子KLP2的表达及多克隆抗体的制备
Expression of Goatpox Virus Virulence Factor KLP2 and Preparation of Polyclonal Antibody
摘要
Abstract
In order to study capripox virus,and prepare polyclonal antibody of its virulence factor KLP2,the prokaryotic expression vector of two structure domains (KLP2-1 and KLP2-2) were built with the method of homologous recombination genes and identified by sequencing.The plasmid were transformed into E.coli BL21 and inducing expressed by IPTG,objective protein were tested by SDS-PAGE and Western blotting,and purified by KCl dyeing rubber cutting.The rabbits were immuned with freund's adjuvant,the prepared antibody were evaluated by Western blotting and ELISA test.The results showed that the expression vector of pET42b-KLP2-1 and pET42b-KLP2-2 were built successfully,and KLP2-1 and KLP2-2 genes were 657 and 984 bp,and expressed about 27 and 38 ku protein,respectively,it was consistent with theories value.The concentrations of rubber cutting purified protein was 1 to 2 mg/mL.The antibody showed obvious specific bands detecting by Western blotting,and its titer were 1 ∶ 512 detecting by ELISA.This result indicated that KLP2-1 and KLP2-2 proteins were expressed successfully,and the corresponding polyclonal antibody were prepared.关键词
羊痘病毒/KLP2基因/蛋白表达/抗体制备Key words
capripox virus/KLP2 gene/protein expression/antibody preparation分类
农业科技引用本文复制引用
李金娜,宋书婷,陈宏伟,刘涛,赵爱云,李有文..山羊痘病毒毒力因子KLP2的表达及多克隆抗体的制备[J].中国畜牧兽医,2017,45(10):2851-2857,7.基金项目
国家自然基金(31360616) （31360616）
塔里木畜牧科技兵团重点实验室开放课题(HS201307) （HS201307）
