禽腺病毒4型鞭毛重组蛋白的原核表达及其免疫效果研究

发布时间：2018-03-17 18:03

  本文选题：禽腺病毒型　切入点：鞭毛基因　出处：《中国兽医杂志》2017年05期 　论文类型：期刊论文

【摘要】：为研究禽腺病毒4型重组鞭毛-fiber2蛋白免疫保护性,将鼠伤寒沙门菌鞭毛蛋白基因与禽腺病毒4型的fiber2基因融合,再将融合基因和单独的fiber2基因分别克隆到原核表达载体p ET-28a,后将构建的p ET-28a-flagellin-fiber2和p ET-28a-fiber2重组载体分别转入大肠杆菌BL21细胞,经IPTG诱导表达,利用SDS-PAGE和Western Blot鉴定表达产物,再将表达产物纯化后免疫21日龄SPF鸡;结果表明,重组菌在37℃培养条件下1.0 mmol/L的IPTG诱导6 h可表达最多的可溶性重组蛋白,Western Blot分析表明,所表达的重组蛋白均能与禽腺病毒血清4型感染阳性血清发生特异性反应,说明所表达蛋白具有良好反应原性,动物试验表明,表达的两种目的蛋白能为鸡提供一定保护力,且低剂量的重组鞭毛-fiber2蛋白产生较好的免疫保护效果。
[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，

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