畜牧与兽医2025，Vol.57Issue(6)：92-97,6.
火鸡疱疹病毒亚克隆对活载体疫苗性能影响的研究
The influence of sub-clone of herpesvirus of turkey on performance of live vector vaccines
摘要
Abstract
This study was to compare the influence of Herpesvirus of Turkey(HVT)sub-clones on the generation of live vector vaccines.Homologous recombination was to insert the MiniF into the non-coding region between UL55 and UL56 of HVT strain Fc126 to generate HVT-MiniF recombinants.After three rounds of plaque picking,three recombinant HVT sub-clones were obtained.Then,using the bacterial artificial chromosome technology,the viral DNA of MiniF recombinant HVT was extracted and was transferred into DH10B competent cells to construct three HVT bacterial artificial chromosomes(BAC).After co-transfection of the expression cassette harboring the infectious bursal disease virus(IBDV)VP2 and the BAC DNA into CEFs(Chicken Embryo Fibroblasts,CEF),the growth kinetics of the vectored HVT re-combinants,the parental strain and the commercial vaccine strain were tested.Finally,serum samples were collected from specific pathogen free(SPF)chickens at 3,4,5 and 6 weeks after immunization with the three recombinant live vectors so as to detect the antibody against IBDV with the method of agar gel precipitation test(AGP).The result showed that we first obtained three purified HVT-MiniF subcloned vi-ruses,followed by three corresponding HVT bacterial artificial chromosomes(BACs)which were termed as HVT BAC-C1,HVT BAC-C2,and HVT BAC-C3,respectively.The RFLP(Restriction Fragment Length Polymorphism,RFLP)assay showed the same major fragments,but with differences between these three BACs.We obtained three recombinants HVT live vectors,namely HVT-VP2-C1,HVT-VP2-C2,and HVT-VP2-C3.The results of the growth kinetics showed that there were significant differences between these strains at 6,24,36 and 72 h(P＜0.05),but no significant differences at 12 and 48 h(P＞0.05).All three HVT recombinants from the three sub-clones induced high AGP antibodies which was still lower than the level of the commercial HVT vectored vaccines.There were obvious differences in AGP antibodies induced by the three recombinants,in which HVT-VP2-C2 produced the highest level of antibody.In summary,this study indi-cated that mutations might be likely to happen in the HVT genome during serial passages,highlighting the importance of selection of vaccine strains with consideration of the effect of sub-clone variations on the efficacy of HVT live-vectored vaccines.关键词
火鸡疱疹病毒/亚克隆/活载体疫苗/细菌人工染色体/免疫效力Key words
turkey herpesvirus/sub-clone/live vector vaccine/bacterial artificial chromosome/immunization efficacy分类
农业科技引用本文复制引用
谢丽华,呙荣兵,黄腾,王继春,王志胜,童玲,栗环环,郑亚婷,刘娅梅,张传健,郭容利,Armando Mario DAMIANI..火鸡疱疹病毒亚克隆对活载体疫苗性能影响的研究[J].畜牧与兽医,2025,57(6):92-97,6.基金项目
广西科技重大专项(桂科AA23062050) （桂科AA23062050）
