中国动物检疫2024，Vol.41Issue(9)：122-128,7.DOI:10.3969/j.issn.1005-944X.2024.09.022
表达H9亚型禽流感病毒HA基因的重组火鸡疱疹病毒构建及应用
Construction and Application of Recombinant Herpesvirus of Turkey Expressing HA Gene of Subtype H9 Avian Influenza Virus
宋新宇 1张茜 1贡鑫 1戚亭 1陈雨 1车艳杰1
作者信息
- 1. 天津瑞普生物技术股份有限公司,天津 300308
- 折叠
摘要
Abstract
In order to construct the recombinant herpesvirus of turkey(HVT)expressing HA gene of H9 subtype avian influenza virus(AIV),based on a Fosmid library continuously covering the whole genome of HVT,the optimized HA gene carrying a Pec complex promoter was inserted between the loci of HVT053 and HVT054(95 318～95 319 nt)in HVT non-essential region for replication using Red/ET homologous recombination technology and Gateway LR cloning system,a recombinant HVT strain(rHVT-H9-HA)expressing HA gene was obtained and characterized.The results showed that the entry plasmid pEntr-HA containing HA gene was successfully constructed.pFosC HA was formed by transferring the expression cassette containing HA gene between No.53 and No.54 gene of the pFosC DEST cosmid by LR reaction;after extracting 5 cosmids,the rescued recombinant virus rHVT-H9-HA strain was obtained by transfecting secondary-generation CEF cells.Consecutive cell passages and animal tests showed that the rHVT-H9-HA strain was with high virus titer,genetic stability and anti-interference against maternal antibodies,and capable of fighting against H9 subtype AIV with long immunization duration;compared to inactivated vaccines,the effect of"one injection against two diseases"was better.A technical support was provided by the study to develop the recombinant HVT live-vector vaccine for H9 subtype AIV.关键词
重组毒株rHVT-H9-HA/火鸡疱疹病毒/HA基因/Red/ET同源重组技术/Gateway LR克隆技术Key words
recombinant rHVT-H9-HA strain/HVT/HA gene/Red/ET homologous recombination technology/Gateway LR cloning system分类
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宋新宇,张茜,贡鑫,戚亭,陈雨,车艳杰..表达H9亚型禽流感病毒HA基因的重组火鸡疱疹病毒构建及应用[J].中国动物检疫,2024,41(9):122-128,7.
