表达IBDV AH1株VP2嵌合法氏囊素三肽基因的重组火鸡疱疹病毒的构建及免疫功能分析

发布时间：2018-05-26 00:58

本文选题：传染性法氏囊病病毒 + VP基因　；参考：《中国兽医科学》2017年09期

【摘要】：为研究传染性法氏囊病(IBD)重组火鸡疱疹病毒(HVT)的构建策略,以HVT FC126基因组非必需区UL44~UL45为插入位点,构建带有大肠杆菌黄嘌呤-鸟嘌呤磷酸转移酶基因序列(gpt)标记的HVT转移载体质粒pUAB-gpt。然后构建表达IBDVAH1株VP2嵌合法氏囊素三肽(BS10)表达盒并克隆入pUAB-gpt,获得转移载体质粒pUAB-Pec-BS10-VP2-Egpt。将该转移载体与HVT DNA共转染鸡胚成纤维细胞(CEF),通过间接免疫荧光试验(IFA)、IBDV抗体夹心ELISA检测、Western-blotting分析,结果显示,获得了表达VP2基因的重组HVT病毒rHVT-Pec-BS10-VP2。将该重组病毒免疫1日龄SPF鸡,免疫后第14天,鸡群可以检测到IBDV抗体。免疫后第21天,脾淋巴细胞增殖活性反应显著。证明IBD重组火鸡疱疹病毒构建成功。
[Abstract]:In order to study the construction strategy of recombinant turkey herpesvirus (HVT) from infectious bursal disease (IBD), the non-essential region UL44~UL45 of HVT FC126 was used as the insertion site. HVT transfer vector plasmid pUAB-gptwith the sequence of Escherichia coli xanthine-guanine phosphotransferase gene was constructed. Then the chimeric expression box of IBDVAH1 strain VP2 was constructed and cloned into pUAB-gpt.The transfer vector plasmid pUAB-Pec-BS10-VP2-Egptwas obtained. The transfer vector was co-transfected with HVT DNA into chicken embryo fibroblasts. The recombinant HVT virus rHVT-Pec-BS10-VP2was obtained by Western blotting analysis by indirect immunofluorescence assay and sandwich ELISA analysis. The recombinant virus was immunized with 1 day old SPF chickens. On the 14th day after immunization, IBDV antibody could be detected in chickens. On the 21st day after immunization, the proliferative activity of spleen lymphocytes was significant. It was proved that IBD recombinant turkey herpesvirus was successfully constructed.
【作者单位】：江苏省农业科学院兽医研究所农业部兽用生物制品工程技术重点实验室国家兽用生物制品工程技术研究中心;
【基金】：国家重点研发计划项目(2016YFD0500800) 国家自然科学基金项目(31272537,31302070,31572504) 江苏省自然科学基金项目(BK20141381,BK20151366) 江苏省农业科技自主创新资金项目[CX(16)1052]
【分类号】：S852.65

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