發(fā)布時(shí)間：2018-06-10 02:59

  本文選題：Ⅰ型鴨肝炎病毒 + VP基因��； 參考：《江蘇農(nóng)業(yè)學(xué)報(bào)》2017年01期

【摘要】：根據(jù)已經(jīng)發(fā)表的Ⅰ型鴨肝炎病毒VP1基因序列,設(shè)計(jì)1對(duì)特異性引物(上下游分別插入Bam H I和Sac I酶切位點(diǎn)),利用PCR技術(shù)擴(kuò)增出VP1基因,經(jīng)Bam H I和Sac I雙酶切后,將其克隆至原核表達(dá)載體p ET-32a上,獲得重組表達(dá)質(zhì)粒p ET-VP1。重組質(zhì)粒轉(zhuǎn)化感受態(tài)細(xì)胞E.coli BL21(DE3),重組蛋白經(jīng)IPTG誘導(dǎo)成功表達(dá)。將重組蛋白切膠免疫6周齡的BALB/c小鼠,3次免疫后采血,制備DHV-Ⅰ的多克隆抗體。SDS-PAGE結(jié)果顯示,成功表達(dá)出的重組蛋白分子量約為46 000。Western-blotting分析結(jié)果表明,該重組蛋白可與His組氨酸鼠單克隆抗體發(fā)生特異性反應(yīng),Western-blotting檢測(cè)抗鼠DHV多抗的結(jié)果顯示,DHV多抗能與目的蛋白發(fā)生特異性反應(yīng)。以上結(jié)果表明,DHV的VP1蛋白在大腸桿菌中成功表達(dá),且制備的DHV多抗能用于VP1蛋白表達(dá)的檢測(cè)。
[Abstract]:According to the published VP1 gene sequence of duck hepatitis I virus, a pair of specific primers were designed. The VP1 gene was amplified by PCR, and was digested by Bam H I and Sac I, respectively. It was cloned into prokaryotic expression vector pET-32a and the recombinant expression plasmid pET-VP1 was obtained. The recombinant plasmid was transformed into E. coli BL21 and DE3, and the recombinant protein was induced by IPTG. The recombinant protein was immunized with 6 weeks old BALB / c mice for 3 times. The polyclonal antibody. SDS-PAGE of DHV- 鈪，

本文編號(hào)：2001744