摘要
[目的]对猪繁殖与呼吸综合症(PRRS)的GPS蛋白进行纯化与免疫活性分析,为建立相应的血清学诊断方法奠定基础。[方法]用构建好的重组表达质粒pGEX-6P-5转化BL21后经IPTG诱导表达。对表达产物进行可溶性分析,再将重组目的蛋白纯化后进行SDS-PAGE鉴定和Western-blot分析,最后用重组抗原进行豚鼠免疫试验。[结果]经层析扫描分析目的蛋白含量占菌体蛋白总量30%左右,纯化后目的蛋白纯度可达80%。经Western-blot及豚鼠免疫试验对纯化蛋白进行分析后证明,表达的蛋白具有良好的反应原性和免疫原性。[结论]该研究为深入研究PRRSVORFs基因及其编码蛋白功能提供了材料,同时也为生产猪繁殖与呼吸综合症病毒基因工程产品奠定了良好基础。
[ Objective ] In this study, we purified and analyzed the immunocompetence of GP5 protein from porcine reproductive and respira- tory syndrome virus, so as to lay a foundation for establishing serological diagnostic method. [ Method] The constructed recombinant expression plasmid pGEX-6P-5 was transformed into BL21, then induced by IPTG for expression. The solubility of expression product was first analyzed; recombinant protein was then isolated and purified for SDS-PAGE and Western blot analysis ; finally its immunity against guinea pig was carried out. [ Result ] Chromatography scanning analysis showed that target protein accounted for about 30% of total protein in bacteria, its purity could reach 80% after purification. Western blot analysis and guinea pig immunity proved that the protein expressed have good reactionogenic- ity and immunogenicity. [ Conclusion ] This study provides substantial material for further researching the ORFs of PRRSV and the function corresponding protein encoded, meanwhile lays a good foundation for preparing genetic vaccine for PRRSV.
出处
《安徽农业科学》
CAS
北大核心
2009年第21期9982-9983,9994,共3页
Journal of Anhui Agricultural Sciences
基金
国家"十一五"高技术研究发展计划(863)项目(2006AA-10A207)
关键词
猪繁殖与呼吸综合征病毒
GP5
表达
纯化
免疫实验
Porcine reproductive and respiratory syndrome virus （ PRRSV ）
GP5 protein
Expression
Purification
Immunological experiment
