非洲猪瘟病毒p30蛋白的原核表达及其多克隆抗体制备  

EXPRESSION OF P30 PROTEIN OF AFRICAN SWINE FEVER VIRUS IN PROKARYOTIC CELLS AND PREPARATION OF POLYCLONAL ANTIBODIES

作　　者：黄剑 徐晶晶[2] 程雪飞 李国新[2] 高飞[2] 童光志 HUANG Jian;XU Jing-jing;CHENG Xue-fei;LI Guo-xin;GAO Fei;TONG Guang-zhi(College of Animal Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)

机构地区：[1]福建农林大学动物科学学院,福州350002 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2020年第5期37-41,共5页Chinese Journal of Animal Infectious Diseases

基　　金：广东省重点领域研发计划项目(2019B020211003);国家自然科学基金专项项目(31941017)。

摘　　要：本研究利用PCR方法扩增非洲猪瘟病毒(ASFV)SY18株的结构蛋白CP204L基因,并将其克隆至原核表达载体pCold I,构建重组表达质粒pCold I-p30,并将其转化至E.coli BL21(DE3)感受态细胞,经异丙基硫代半乳糖苷(IPTG)诱导表达,SDS-PAGE显示,p30蛋白在大肠杆菌大量表达。获得的p30经纯化免疫小鼠,4次免疫后,采取其血清经过间接免疫荧光和Western blot检测结果表明,获得的多克隆抗体具有明显的特异性。该研究获得的针对p30的多克隆抗体为ASFV早期诊断试及p30蛋白结构功能研究奠定了基础。The CP204L gene(SY18)of African swine fever virus synthesized was amplified by PCR and inserted into the multiple clone sites of the pCold I vector.The recombinant plasmid was transformed into BL21(DE3)competent cells to express the p30 gene with induction of IPTG.SDS-PAGE analysis showed that the p30 protein was obviously expressed.Mice were immunized 4 times at a 2 weeks’interval with the purified p30 protein to prepare antiserum samples.The polyclonal antibodies had significant specificity as shown in indirect immunofluorescence and Western blot.The availability of the polyclonal antibodies against P30 protein laid a foundation for early diagnosis of ASFV and research on its structure and function.

关 键 词：非洲猪瘟病毒 P30基因 原核表达 多克隆抗体 

分 类 号：S852.651[农业科学—基础兽医学]