新城疫病毒F蛋白在水稻中的表达及检测  

Expression and Detection of NDV-F Protein in Rice Endosperm

作　　者：许倩茹[1] 张二芹[1] 郭军庆[2] 杨继飞[2] 刘运超[2] 王爱萍[3] 王丽[2] 万博[1] 汪磊[1] 蒋大伟[1] 邓瑞广[2] 张改平[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州450002 [2]河南省农业科学院动物免疫学重点实验室农业部动物免疫学重点实验室河南省动物免疫学重点实验室,郑州450002 [3]郑州大学生命科学学院,郑州450001

出　　处：《畜牧兽医学报》2017年第6期1167-1172,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：兽用蛋白功能性转基因水稻新品种培育(2016ZX08001006);公益性行业(农业)科研专项经费项目(201303033)

摘　　要：旨在研究水稻表达新城疫病毒F蛋白,制备亚单位疫苗。根据GenBank中的新城疫病毒F基因序列以及水稻偏爱性密码子筛选、优化并合成F基因,通过两次载体构建,先使目的基因连在中间载体pMP3上,再连接到pCAMBIA1300植物载体上,通过农杆菌导入水稻愈伤组织中,经过潮霉素抗性筛选,获得表达植株。通过QPCR的方法筛选获得高表达的纯合子株系5株。新城疫抗原试纸条检测和Western blot检测确定F蛋白具有良好的反应原性。本试验为家禽传染病的植物性水稻疫苗研究奠定了基础。To develop rice-derived subunit vaccine of Newcastle disease （ND）, NDV F gene was screened, designed and synthesis after clone and construct recombinant plasmid pCAMBIA1300-F according to rice preference codon optimization. By constructing of vector twice, we made the tar- get gene connect with vector pMP3, then transferred the gene to the vector pCAMBIA1300, fi- nally Agrobacterium tumefaciens would be a mediate witch lead the gene to rice callus. The expressed rice seeds were got after screening by hygromycin. To obtain high expression of homo- zygous strain, the real time PCR were used to select 5 homozygous lines. The test of NDV-F an- tigen strip and Western blot indicated that the recombinant protein could react with good antigen- ic activity. This research would lay the foundation of the rice vaccine research.

关 键 词：新城疫病毒 F基因 植物表达载体 QPCR 纯合子 

分 类 号：S852.659.5[农业科学—基础兽医学] Q789[农业科学—兽医学]