本文選題：禽腺病毒型　切入點(diǎn)：鞭毛基因　出處：《中國(guó)獸醫(yī)雜志》2017年05期 　論文類型：期刊論文

【摘要】：為研究禽腺病毒4型重組鞭毛-fiber2蛋白免疫保護(hù)性,將鼠傷寒沙門菌鞭毛蛋白基因與禽腺病毒4型的fiber2基因融合,再將融合基因和單獨(dú)的fiber2基因分別克隆到原核表達(dá)載體p ET-28a,后將構(gòu)建的p ET-28a-flagellin-fiber2和p ET-28a-fiber2重組載體分別轉(zhuǎn)入大腸桿菌BL21細(xì)胞,經(jīng)IPTG誘導(dǎo)表達(dá),利用SDS-PAGE和Western Blot鑒定表達(dá)產(chǎn)物,再將表達(dá)產(chǎn)物純化后免疫21日齡SPF雞;結(jié)果表明,重組菌在37℃培養(yǎng)條件下1.0 mmol/L的IPTG誘導(dǎo)6 h可表達(dá)最多的可溶性重組蛋白,Western Blot分析表明,所表達(dá)的重組蛋白均能與禽腺病毒血清4型感染陽(yáng)性血清發(fā)生特異性反應(yīng),說(shuō)明所表達(dá)蛋白具有良好反應(yīng)原性,動(dòng)物試驗(yàn)表明,表達(dá)的兩種目的蛋白能為雞提供一定保護(hù)力,且低劑量的重組鞭毛-fiber2蛋白產(chǎn)生較好的免疫保護(hù)效果。
[Abstract]:In order to study the immune protection of recombinant flagell-fiber2 protein of avian adenovirus type 4, the flagellin gene of Salmonella typhimurium was fused with the fiber2 gene of avian adenovirus type 4. Then the fusion gene and the single fiber2 gene were cloned into the prokaryotic expression vector pET-28a, and then the recombinant vectors of p ET-28a-flagellin-fiber2 and p ET-28a-fiber2 were transformed into E. coli BL21 cells respectively. The expression products were identified by SDS-PAGE and Western Blot after IPTG induced expression. The expression product was purified and immunized with 21-day-old SPF chicken. The results showed that the recombinant strain could express the most soluble recombinant protein after 6 h induction with 1.0 mmol/L IPTG at 37 鈩，

本文編號(hào)：1625853