牛传染性鼻气管炎病毒(gN-TK-gG-)三基因缺失突变株的构建  

Construction of a triple gene deleted mutant BoHV-1 gN^-/TK^-/gG^-

作　　者：李程 陈颖钰[2,3] 索朗斯珠[1] 郭爱珍 LI Cheng;CHEN Ying-yu;SUOLANG Si-zhu;GUO Ai-zhen(College of Animal Science,Tibet Agricultural ＆ Animal Husbandry University,Linzhi,Tibet 860000,China;College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China;State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]西藏农牧学院动物科学学院,西藏林芝860000 [2]华中农业大学动物医学院,湖北武汉430070 [3]华中农业大学农业微生物学国家重点实验室,湖北武汉430070

出　　处：《中国兽医学报》2018年第7期1276-1281,共6页Chinese Journal of Veterinary Science

基　　金：高校博士点基金资助项目(20130146110003);现代农业(肉牛/牦牛)产业技术体系专项资金资助项目(CARS-38);国家重点研发计划资助项目(2016YFD0500906)

摘　　要：牛传染性鼻气管炎是由牛疱疹病毒I型（BoHV-1）引起的一种牛的传染病，可导致严重的经济损失。疫苗接种是用于防控该病的重要手段，但现有疫苗在多个方面都有改进的余地。本试验利用pBoHV-1 BAC载体和4次Red E／T重组，依次构建了BoHV-1 gN^-（CT^-）、BoHV-1 gN^-（CT^-30-32^-）、BoHV-1 gN^-TK^-和BoHV-1 gN TK gG^-基因缺失突变株，其中gN缺失了胞外区30-32位氨基酸、胞质尾区CT端，TK基因缺失了1079bp，gG基因缺失了1335bp。pBoHV-1 gN^-TK^-gG^-经磷酸钙法转染MDBK细胞后，出现细胞病变，成功拯救出三基因缺失病毒株BoHV-1 gN TK^-gG^-。研究结果为牛传染性鼻气管炎新型基因缺失标记疫苗的开发奠定了基础。Bovine infectious rhinotracheitis is a bovine infectious disease caused by bovine herpesvirus type I（BoHV-1）,which can lead to serious economic losses. Vaccination is an important mean to prevent and control this disease. However,the current vaccines remain to be improved in many ways. In this study,BoHV-1 gN^- （CT null） ,BoHV-1 gN^- （CT/30-32 null） ,BoHV-1 gN TK and BoHV-1 gN^-TK gG^- were sequentially constructed by using pBoHV-1 BAC vector and four rounds of Red E/T recombination. The gN was deleted of extracellular domain（including 9 bp）, the part of cytoplasmic tail（CT）（covering 7 bp）,while TK gene was deleted of 1 079 bp,and gG gene of 1 335 bp. The MDBK cells were transfected by the final recombinant plasmid pBoHV-1 gN^- TK^- gG^- with calcium phosphate method and the virus replication was observed in the cells shown by green fluorescence indicating that the viral mutant BoHV-1 gN^-TK^-gG^- was success- fully rescued. This result provides the basis for further development of novel vaccine with triple gene deleted marker against bovine infectious rhinotracheitis.

关 键 词：牛传染性鼻气管炎 牛疱疹病毒I型 基因缺失疫苗 gN TK GG 

分 类 号：S852.65[农业科学—基础兽医学]