摘要
利用生物信息学软件对减蛋综合征病毒(EDSV)AV-127纤维蛋白进行抗原特性分析,人工合成融合基因Knobs,将其克隆到原核表达载体pET28a的多克隆位点,构建原核表达载体pET28a-Knobs,将其转化到感受态细胞Rosetta(DE3)pLysS中,经30℃、1.0 mmol.L-1IPTG诱导表达,SDS-PAGE分析超声裂解后的上清和沉淀。结果显示:目的蛋白以包涵体形式存在,蛋白分子质量约为26.0 ku;Western blot结果表明目的蛋白具有较好的抗原活性。本试验为进一步研究EDSV抗原结构域的免疫原性以及减蛋综合征基因工程疫苗的开发奠定了基础。
Antigenicity of egg drop syndrome virus （EDSV） strain AV-127 fiber protein was analysed using bioinformatics software. The gene fragment knobs coding for the antigenic structural domains of EDSV fiber protein was synthesized, and then cloned into the polycloning sites of pET28a vector. The recombinant prokaryotic expression plasmid was constructed, and named pET28a-Knobs. The recombinant plasmid was transformed into competent cells Rosetta （ DE3 ） pLysS and induced for expression by 1.0 mmol/L of IPTG. The SDS-PAGE analysis showed that the expressed fusion protein with a molecular weight of 63.5 ku existed in the inclusion body. The Western blot analysis showed that the protein had good antigenicity. This study will apply a basis for the further antigenicity analysis of these antigenic structural domains and gene engineering vaccines of EDSV.
出处
《畜牧与兽医》
北大核心
2012年第2期22-25,共4页
Animal Husbandry & Veterinary Medicine
基金
江西省科技厅(2011BDH80035
2010BNA08000)
江西省科学院(2010-YGJ-01
2010-YGJ-02)
关键词
减蛋综合征病毒
结构域
原核表达
抗原
egg drop syndrome virus （EDSV）
structural domains
prokaryotic expression
antigenicity
