中国兽医科学2012，Vol.42Issue(3)：268-274,7.
鸡朊蛋白的原核表达及其抗血清的制备
Prokaryotic expression of chicken prion protein and preparation of its antiserum
摘要
Abstract
To study the structure and function of chicken prion protein（ChPrP）,three recombinant expression vectors（pGEX-4T-ChPrP1,pGEX-4T-ChPrP2 and pGEX-4T-ChPrP3） were constructed,and were transformed into Escherichia coli BL21（DE3）,respectively.The fermentation conditions including time and IPTG concentration were optimized.Fusion protein GST-ChPrP（25-247） of pGEX-ChPrP1 was purified by GST-trap FF affinity chromatography and was used to immunize New Zealand male rabbits.The results showed that an efficient prokaryotic expression system of pGEX-4T-ChPrP1,pGEX-4T-ChPrP2,and pGEX-4T-ChPrP3 was successfully constructed.SDS-PAGE analysis showed that three IPTG-induced fusion proteins GST-ChPrP（25-247）,GST-ChPrP（96-247）,and GST-ChPrP（174-247） were expressed as expected with the molecular weight of about 50,42,and 32 ku,respectively.Under optimal fermentation conditions,10 mg fusion protein GST-ChPrP（25-247） could obtain by GST-trap FF affinity chromatography in one liter bacilli of pGEX-ChPrP1.The results of agar diffusion test showed that prepared polyclonal antiserum has a high antibody titer（1∶32）.Western-blot analysis proved that the three expressed fusion protein and the extration of chicken brain tissue can specifically reacted with the antiserum respectively, demonstrating that the recombinant chicken prion protein possesses good antigenicity and the same antigenic components in brain tissue.关键词
鸡/朊蛋白/原核表达/抗血清/抗原性Key words
chicken/prion protein/prokaryotic expression/antiserum/antigenicity分类
农业科技引用本文复制引用
罗鹏,吴润,刁小龙,管宏伟,张莉..鸡朊蛋白的原核表达及其抗血清的制备[J].中国兽医科学,2012,42(3):268-274,7.基金项目
国家自然科学基金项目 （）
甘肃省自然科学研究基金计划项目 （）
家畜疫病病原生物学国家重点实验室开放基金项目 （）
