本文關(guān)鍵詞： 豬繁殖與呼吸綜合征 NSP 免疫原性　出處：《中國畜牧獸醫(yī)》2017年06期 　論文類型：期刊論文

【摘要】：試驗(yàn)旨在表達(dá)純化豬繁殖與呼吸綜合征病毒NSP2重組蛋白。利用PCR技術(shù)擴(kuò)增獲得大小為750bp的NSP2 226-475位氨基酸編碼序列,然后將該序列亞克隆到pET-28a(+)表達(dá)載體上,通過pET-28a(+)表達(dá)載體在大腸桿菌Rosetta-gami 2(DE3)pLys中得以表達(dá),獲得大小為29ku的重組蛋白,對(duì)重組蛋白誘導(dǎo)時(shí)間進(jìn)行優(yōu)化,SDS-PAGE顯示重組蛋白在37℃經(jīng)1mmol/L IPTG誘導(dǎo)8h時(shí)表達(dá)量最大,免疫印跡結(jié)果證實(shí)表達(dá)的蛋白能被PRRSV陽性血清識(shí)別,具有良好的免疫原性。本研究成功表達(dá)純化了NSP2重組蛋白,為目標(biāo)蛋白作為包被抗原建立檢測PRRSV血清抗體的ELISA方法奠定了基礎(chǔ)。
[Abstract]:The aim of this study was to express and purify the recombinant NSP2 protein of porcine reproductive and respiratory syndrome virus (RRRSV). A 750bp NSP2 226-475 amino acid encoding sequence was amplified by PCR technique and subcloned into pET-28a() expression vector. The recombinant protein of 29ku was obtained by expressing pET-28a () expression vector in E. coli Rosetta-gami 2DE3DE3pLys. The optimal induction time of the recombinant protein was optimized by SDS-PAGE. The expression of the recombinant protein was the highest at 37 鈩，

本文編號(hào)：1518995