本文選題：水泡性口炎病毒(VSV) + N蛋白��； 參考：《中國畜牧獸醫(yī)》2017年09期

【摘要】：本研究旨在克隆和表達(dá)水泡性口炎病毒(vesicular stomatitis virus,VSV)特異性抗原N蛋白,進(jìn)而純化并分析其免疫原性。根據(jù)GenBank中已發(fā)表的VSV基因組N基因序列,分別合成VSV兩種不同血清型的N基因,經(jīng)序列對比分析后,設(shè)計(jì)合成1對特異性引物,PCR擴(kuò)增獲得約1 300bp的N基因片段,將目的片段亞克隆至pColdⅠ原核表達(dá)載體中,經(jīng)IPTG誘導(dǎo)表達(dá)后,采用Ni-NTA樹脂親和層析法純化重組N蛋白。SDS-PAGE分析表明,N基因在大腸桿菌中得到表達(dá),蛋白大小約為50ku;Western blotting檢測結(jié)果表明,該重組蛋白與VSV多克隆抗體發(fā)生特異性反應(yīng)。本試驗(yàn)成功構(gòu)建了VSV-IND和VSV-NJ的原核表達(dá)載體,實(shí)現(xiàn)了N蛋白在大腸桿菌中的可溶性表達(dá),純化后的重組蛋白具有良好的免疫原性。
[Abstract]:The aim of this study was to clone and express the specific antigen N protein of vesicular stomatitis virus, and then purify and analyze its immunogenicity. According to the N gene sequence of VSV genome published in GenBank, the N gene of two different serotypes of VSV was synthesized. After sequence comparison and analysis, a pair of specific primers were designed and synthesized to amplify the N gene fragment of about 1 300bp. The target fragment was subcloned into the prokaryotic expression vector of pCold 鈪，

本文編號：1794802