牛疱疹病毒1型糖蛋白E(gE)可溶性表达及其反应原性分析  

Soluble expression and reactogenicity analysis of glycoprotein E(gE) of bovine herpesvirus type 1

作　　者：杨晨 孙续 丁学智[1] 杨峰[1] YANG Chen;SUN Xu;DING Xuezhi;YANG Feng(Key Lab of New Veterinary Durg Project in Gansu Province,Key Lab of Veterinary Pharmaceutics Discovery,Ministry of Agricultural and Rural Affairs,Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS,Lanzhou Gansu 730050,China)

机构地区：[1]中国农业科学院兰州畜牧与兽药研究所,农业农村部兽用药物创制重点实验室,甘肃省新兽药工程重点实验室,甘肃兰州730050

出　　处：《中兽医医药杂志》2023年第3期17-21,共5页Journal of Traditional Chinese Veterinary Medicine

基　　金：甘肃省重点研发计划项目(21YF5NA141);国家重点研发计划项目(2022YFD1302101)。

摘　　要：为探索牛传染性鼻气管炎诊断新技术,本研究以GenBank公布的牛疱疹病毒1型(BHV1,NC_001847.1)基因序列为参考,预测并优化了BHV1的gE蛋白胞外区密码子序列。优化后的序列长度为1 275 bp,5’端和3’端分别添加限制性酶切位点SacⅠ和HindⅢ后克隆至载体pUC19,进而构建原核表达载体pET-SUMO-gE,将其转入大肠杆菌BL21-CodonPlus(DE3)-RIL中表达重组蛋白,使用NiNTA亲和柱纯化重组蛋白。SDS-PAGE和Western blot检测结果证明,获得的牛疱疹病毒1型gE胞外区重组蛋白大小约为70 kDa,有良好的水溶性和反应原性。该研究结果为gE特异性单克隆抗体的制备及后续诊断试剂盒的研发奠定了基础。In order to explore a new diagnostic technique for bovine infectious rhinotracheitis,in this study,the extracellular domain sequence of gE protein in bovine herpesvirus type 1(BHV1) was predicted and optimized according to the BHV1(NC_001847.1) gene published by GenBank as the reference sequence.The optimized sequence length was1 275 bp,restriction sites SacⅠ and Hind Ⅲ were added to the 5' end and 3' end,respectively,and cloned into vector pUC19.The prokaryotic expression vector pET-SUMO-gE was constructed and transferred into Escherichia coli BL21-CodonPlus(DE3)-RIL to express the recombinant protein.The recombinant protein was purified using Ni-NTA affinity columns.The results of SDS-PAGE and Western blot showed that the recombinant protein of gE extracellular region of BHV1 was about 70 kDa with good water solubility and reactivity.The results of this study lay a foundation for the preparation of gE specific monoclonal antibody and the development of subsequent diagnostic kits.

关 键 词：牛传染性鼻气管炎 牛疱疹病毒1型 gE蛋白 原核表达 载体构建 

分 类 号：S858.23[农业科学—临床兽医学]