兔出血症病毒VP60截短基因原核表达及诱导条件优化  

Optimization of Induction Conditions for Prokaryotic Expression of Rabbit Hemorrhagic Disease Virus VP60 Truncated Gene

作　　者：郑庆礼 许高琳 肖春龙 王全溪[3,4] ZHENG Qingli;XU Gaolin;XIAO Chunlong;WANG Quanxi(College of Animal Science and Technology,Fujian Agricultural Vocational and Technical College,Fuzhou 350007;Fujian Chunlong Agriculture&Animal Husbandry Technology Company Limited,Minhou 350100;Fujian Key Laboratory of Veterinary Chinese Medicine and Animal Health,Fuzhou 350002;Fujian Agriculture and Forestry University,Fuzhou 350002)

机构地区：[1]福建农业职业技术学院动物科技学院,福州350007 [2]福建省兽医中药与动物保健重点实验室,闽侯350100 [3]福建农林大学,福州350002 [4]福建省春龙农牧科技有限公司,福州350002

出　　处：《中国养兔》2023年第2期8-11,共4页Chinese Journal of Rabbit Farming

基　　金：自动化密闭式兔舍疾病防控关键技术的研究与示范(福建省星火计划)(2020S0005);福建省中青年教师教育科研项目(科技类)(JAT220609)。

摘　　要：兔瘟是由兔出血症病毒(RHDV)引起的传染病,给肉兔养殖产业带来巨大损失。根据RHDV VP60基因序列进行抗原肽分析,选择该基因主要抗原肽段后,设计一对特异性引物进行PCR扩增,并插入pET32a(+)载体中构建重组质粒。经PCR鉴定、酶切鉴定和测序,结果表明p ET32a(+)-VP60截短基因质粒构建成功。对RHDV-VP60截短基因蛋白原核表达系统筛选诱导条件,结果表明:IPTG浓度为0.8 mmoL/L,诱导时间为8 h,诱导温度为37℃,表达量最高。研究为后续兔瘟亚单位疫苗和诊断试剂的研究奠定基础。Rabbit plague is an infectious disease caused by rabbit hemorrhagic disease virus(RHDV),which brings huge losses to the rabbit breeding industry.According to the antigenic peptide analysis of RHDV VP60 gene sequence,after selecting the main antigenic peptide segment of the gene,a pair of specific primers were designed for PCR amplification and inserted into pET32a(+)vector to construct the recombinant plasmid.The results of PCR,restriction enzyme digestion and sequencing showed that the pET32a(+)-VP60 truncated gene plasmid was successfully constructed.The results showed that the expression level was the highest,which the concentration of IPTG was 0.8 mmoL/L,the induction time was 8 h,the induction temperature was 37°C.This study lays a foundation for the follow-up study of rabbit plague subunit vaccine and diagnostic reagents.

关 键 词：兔出血症病毒 VP60基因 原核表达 诱导条件 

分 类 号：S829.1[农业科学—畜牧学]