表达FAdV-8b Fiber蛋白重组FAdV-4的构建与鉴定  

Construction and identification of recombinant fowl adenovirus 4 expressing the fiber gene of fowl adenovirus 8b

作　　者：王白玉 黄庆 乔麒龙 杨盼盼 李永涛[1] 王增 杨霞[1] 苗玉和 赵军[1] WANG Baiyu;HUANG Qing;QIAO Qilong;YANG Panpan;LI Yongtao;WANG Zeng;YANG Xia;MIAO Yuhe;ZHAO Jun(College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China;Fujian Shengwei Biotech Co.,Ltd.,Nanping,Fujian 354100,China)

机构地区：[1]河南农业大学动物医学院,河南郑州450046 [2]福建圣维生物技术有限公司,福建南平354100

出　　处：《中国兽医学报》2022年第2期230-236,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31772771)。

摘　　要：为了构建能够同时防控鸡肝炎-心包积液综合征(HHS)和包涵体肝炎(IBH)的新型疫苗,本研究利用禽腺病毒4型(FAdV-4)中国流行株CH/HNJZ/2015的反向遗传操作技术,将禽腺病毒8b型(FAdV-8b)中国流行株的全长Fiber基因插入至FAdV-4基因组中1 966 bp自然缺失区,构建感染性克隆并拯救出重组病毒rHNJZ-Fiber/FAdV8b,利用PCR、Western blot和间接免疫荧光试验对重组病毒rHNJZ-Fiber/FAdV8b进行鉴定。结果表明,FAdV-8b的Fiber基因得以准确插入,Fiber蛋白正确并稳定表达,重组病毒rHNJZ-Fiber/FAdV8b具有良好的遗传稳定性,在细胞中具有与亲本毒株相似的高滴度生长特性和复制动态。本研究构建的表达FAdV-8b中国流行株Fiber基因的重组FAdV-4将为同时研发HHS和IBH的有效疫苗提供新的思路。A recombinant fowl adenovirus serotype 4(FAdV-4) expressing the Fiber gene of fowl adenovirus serotype 8 b(FAdV-8 b) was constructed in this study to establish a basis for the development of a novel vaccine that protect chickens against both hepatitis-hydropericardium syndrome(HHS) caused by FAdV-4 and inclusion body hepatitis(IBH) caused by FAdV-8 b.The full sequence of the fiber gene of FAdV-8 b epidemic strain was inserted into the 1 966 bp deletion area of FAdV-4 Chinese epidemic strain CH/HNJZ/2015 using a reverse genetic technique of FAdV-4.The rescued recombinant virus rHNJZ-Fiber/FAdV-8 b was identified by PCR,sequencing, Western blot, and indirect immunofluorescence assay.The results indicated that the Fiber gene was successfully inserted into the 1 966-bp deletion area and the Fiber protein was correctly expressed.The recombinant virus rHNJZ-Fiber/FAdV-8 b is genetically stable and has similar growth characteristics to the parent strain CH/HNJZ/2015.The recombinant virus rHNJZ-Fiber/FAdV-8 b will be a cost-effective candidate vaccine for the efficient control and prevention of HHS and IBH.

关 键 词：禽腺病毒4型 禽腺病毒8b型 Fiber蛋白 肝炎-心包积液综合征 包涵体肝炎 新型疫苗 

分 类 号：S852.65[农业科学—基础兽医学]