利用杆状病毒表达系统表达鸡传染性法氏囊病病毒VP2蛋白

发布时间：2018-01-24 18:49

本文关键词： 传染性法氏囊病毒杆状病毒表达载体 VP蛋白　出处：《畜牧与兽医》2017年08期 　论文类型：期刊论文

【摘要】：从GenBank下载传染性法氏囊病病毒(IBDV)VP2基因序列,根据昆虫细胞偏爱密码子对IBDV VP2基因进行密码子优化,化学合成优化后的VP2基因序列,然后构建杆状病毒表达载体pTri Ex-4-VP2,构建成功的重组杆状病毒pTri Ex-4-VP2转染sf9细胞,制备病毒原种,采用SDS-PAGE和Western blot方法鉴定VP2的免疫原性。结果显示:细胞感染Bac-VP2后,其细胞裂解蛋白在58 ku附近有1个条带,且该条带与IBD阳性血清发生特异性反应;间接免疫荧光试验(IFA)结果显示VP2基因能在Sf9细胞中表达;动物攻毒保护试验中,2次免疫重组VP2蛋白后能诱导14日龄SPF鸡产生对IBDV强毒攻击,保护率为60%。本研究为进一步研究IBDV VP2基因工程疫苗提供了物质基础。
[Abstract]:The sequence of VP2 gene of infectious bursal disease virus (IBDV) was downloaded from GenBank, and the codon of IBDV VP2 gene was optimized according to insect cell preference codon. The optimized VP2 gene sequence was synthesized by chemical synthesis, and then the baculovirus expression vector pTri Ex-4-VP2 was constructed. The recombinant baculovirus pTri Ex-4-VP2 was constructed and transfected into sf9 cells. SDS-PAGE and Western blot were used to identify the immunogenicity of VP2. The cleavage protein had one band near 58 ku, and it reacted specifically with IBD positive serum. The results of indirect immunofluorescence assay (IFA) showed that VP2 gene could be expressed in Sf9 cells. The recombinant VP2 protein was immunized twice in the animal toxicity protection test, and it could induce 14 day old SPF chickens to attack IBDV strongly. The protective rate is 60%. This study provides a material basis for further study of IBDV VP2 gene engineering vaccine.
【作者单位】：广东温氏大华农生物科技有限公司;华南农业大学兽医学院;
【基金】：省应用型科技研发专项资金项目(2015B020230011)
【分类号】：S852.65
【正文快照】： 鸡传染性法氏囊病(infectious bursal disease,IBD)是由传染性法氏囊病毒(infectious bursaldisease virus,IBDV)侵害引起的一种急性、高度接触性传染病,主要侵害雏鸡和青年鸡的法氏囊[1-2]。该病不仅造成鸡只死亡还导致免疫抑制,是目前危害世界养禽业的主要传染病之一[3]。传

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