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.
作者
王白玉
黄庆
乔麒龙
杨盼盼
李永涛
王增
杨霞
苗玉和
赵军
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)
出处
《中国兽医学报》
CAS
CSCD
北大核心
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)。
