新城疫病毒F蛋白在水稻中的表达及检测

doi:10.11843/j.issn.0366-6964.2017.06.022

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (6): 1167-1172.doi: 10.11843/j.issn.0366-6964.2017.06.022

• 研究简报 • 上一篇

新城疫病毒F蛋白在水稻中的表达及检测

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*

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

收稿日期:2017-01-10 出版日期:2017-06-23 发布日期:2017-06-23
通讯作者: 张改平(1960-),男,河南内黄人,教授,主要从事动物免疫学研究,E-mail:zhanggaiping2003@163.com
作者简介:许倩茹(1991-),女,河南郑州人,硕士生,主要从事动物免疫学研究,E-mail:xuqianru2017@163.com
基金资助:
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）

Expression and Detection of NDV-F Protein in Rice Endosperm

XU Qian-ru¹, ZHANG Er-qin¹, GUO Jun-qing², YANG Ji-fei², LIU Yun-chao², WANG Ai-ping³, WANG Li², WAN Bo¹, WANG Lei¹, JIANG Da-wei¹, DENG Rui-guang², ZHANG Gai-ping^1*

1. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China;
2. Key Laboratory of Animal Immunology of the Ministry of Agriculture, Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China;
3. College of Life Science, Zhengzhou University, Zhengzhou 450001, China

Received:2017-01-10 Online:2017-06-23 Published:2017-06-23

摘要/Abstract

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。

Abstract:

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 target gene connect with vector pMP3, then transferred the gene to the vector pCAMBIA1300, finally 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 homozygous strain, the real time PCR were used to select 5 homozygous lines. The test of NDV-F antigen strip and Western blot indicated that the recombinant protein could react with good antigenic activity. This research would lay the foundation of the rice vaccine research.

中图分类号:

许倩茹, 张二芹, 郭军庆, 杨继飞, 刘运超, 王爱萍, 王丽, 万博, 汪磊, 蒋大伟, 邓瑞广, 张改平. 新城疫病毒F蛋白在水稻中的表达及检测[J]. 畜牧兽医学报, 2017, 48(6): 1167-1172.

XU Qian-ru, ZHANG Er-qin, GUO Jun-qing, YANG Ji-fei, LIU Yun-chao, WANG Ai-ping, WANG Li, WAN Bo, WANG Lei, JIANG Da-wei, DENG Rui-guang, ZHANG Gai-ping. Expression and Detection of NDV-F Protein in Rice Endosperm[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(6): 1167-1172.

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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

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