利用玉米作为生物反应器表达禽呼肠孤病毒σC抗原蛋白  

Expression of σC Core Neutralizing Epitope Protein of Avian Reovirus in Maize Bioreactor

作　　者：王盛 谢芝勋 万丽军 谢丽基 罗思思 范晴 张艳芳 曾婷婷 黄娇玲 张民秀 谢志勤 邓显文 WANG Sheng;XIE Zhixun;WAN Lijun;XIE Liji;LUO Sisi;FAN Qing;ZHANG Yanfang;ZENG Tingting;HUANG Jiaoling;ZHANG Minxiu;XIE Zhiqin;DENG Xianwen(Guangxi Veterinary Research Institute,Guangxi Key Laboratory of Veterinary Biotechnology,Nangning 530001,China)

机构地区：[1]广西壮族自治区兽医研究所广西兽医生物技术重点实验室,南宁530001

出　　处：《中国动物传染病学报》2022年第3期166-172,共7页Chinese Journal of Animal Infectious Diseases

基　　金：广西自然科学基金项目(2017GXNSFBA198140);广西自然科学基金项目(2018GXNSFAA138106);广西科技基地和人才专项(桂科AD17195083);广西八桂学者专项(2019A50);国家“万人计划”领军人才专项(W02060083)。

摘　　要：本研究采用玉米作为生物反应器表达禽呼肠孤病毒(ARV)σC抗原蛋白。首先,根据NCBI数据库中ARV σC基因的序列及玉米中高表达蛋白密码子的偏好性,用于软件分析优化σC基因序列,并人工合成σC基因;将σC基因克隆到植物表达载体pRI201,将重组质粒转化农杆菌GV3101,筛选获得阳性菌株。随后通过农杆菌介导法,将σC基因转化玉米愈伤组织,筛选获得转基因玉米植株。PCR、RT-PCR鉴定结果表明,ARV σC基因成功的整合到玉米基因组中,并能进行正确的转录和翻译。Western blot结果显示,玉米中表达的σC蛋白可以被ARV阳性血清识别,具有良好的反应原性。本研究为ARV新型植物口服疫苗的研究奠定了基础。The σC core neutralizing epitope of Avian reovirus (ARV) was expressed in maize (Zea may L.) bioreactor in order to explore the possibility of developing a novel plant oral vaccine.Firstly,according to the σC gene sequence published on NCBI and the codon bias of maize,optimizing σC gene codon sequence was synthesized for insertion into plant expression vector pRI201 and transformation into Agrobacterium strain of GV3101.The σC gene was transformed into maize via agrobacterium for regeneration of maize plants.The result showed that the σC gene was integrated into the maize genome as tested by PCR and RT-PCR and the expressed σC protein had good biological activity as demonstrated in Western blot.Thus,this experiment laid a foundation for developing an ARV plant oral vaccine.

关 键 词：ARV σC基因 转基因玉米 口服疫苗 

分 类 号：S852.65[农业科学—基础兽医学]