HPV45-L1和HPV58-L1蛋白植物表达载体的构建及水稻转基因植株的鉴定

doi:10.7668/hbnxb.2017.02.009

华北农学报 ›› 2017, Vol. 32 ›› Issue (2): 55-60. doi: 10.7668/hbnxb.2017.02.009

所属专题： 水稻；生物技术；

HPV45-L1和HPV58-L1蛋白植物表达载体的构建及水稻转基因植株的鉴定

王聚财¹, 刘运超², 陈玉梅^2,3, 孟凤丽^2,3, 王爱萍³, 张二芹¹, 许倩茹¹, 邓瑞广², 张改平¹

1. 河南农业大学牧医工程学院, 河南郑州 450002;
2. 河南省农业科学院河南省动物免疫学重点实验室, 农业部动物免疫学重点实验室, 河南郑州 450002;
3. 郑州大学生命科学学院, 河南郑州 450001

收稿日期:2016-11-08 出版日期:2017-04-28
通讯作者: 张改平(1960-),男,河南内黄人,院士,博士生导师,主要从事动物免疫学及动物病毒分子致病机制研究。
作者简介:王聚财(1990-),女,河南洛阳人,硕士,主要从事疫病与疫苗研究。
基金资助:
河南省高校科技创新团队和创新人才支持计划（14HASTIT027）

Construction of HPV45-L1 and HPV58-L1 Plant Expression Vectors and Identification of Transgenic Rice Plants

WANG Jucai¹, LIU Yunchao², CHEN Yumei^2,3, MENG Fengli^2,3, WANG Aiping³, ZHANG Erqin¹, XU Qianru¹, DENG Ruiguang², ZHANG Gaiping¹

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

Received:2016-11-08 Published:2017-04-28

摘要/Abstract

摘要： 利用水稻胚乳细胞生物反应器，构建含有HPV45-L1和HPV58-L1基因的重组植物表达载体，为HPV-45L1和HPV58-L1蛋白的表达提供一种新的高效、低廉的表达方式。利用PCR技术克隆人乳头瘤病毒HPV45-L1和HPV58-L1蛋白编码基因，将其重组于中间载体pMP3和植物表达载体pCAMBIA-1300中，构建含HPV45-L1和HPV58-L1基因的植物表达载体pCAMBIA-1300-pMP3-HPV45-L1、pCAMBIA-1300-pMP3-HPV58-L1，随后采用根癌农杆菌侵染水稻愈伤组织介导的水稻遗传转化，获得HPV-L1转基因水稻植株。PCR检测结果表明，HPV45-L1和HPV58-L1基因已整合到水稻基因组中，说明已成功构建水稻胚乳特异性表达载体。

关键词: 人乳头瘤病毒, 转基因水稻, 水稻胚乳特异性启动子, 根癌农杆菌

Abstract: The rice seeds were used as the cell bioreactor to construct the recombinant plant expression vector containing HPV45-L1 and HPV58-L1 genes to provide a new efficient and inexpensive method for HPV45-L1 and HPV58-L1 protein expression.The HPV45-L1 and HPV58-L1 coding sequence (ORF)was amplified by PCR and then subcloned into middle vector pMP3 and plant expression vector pCAMBIA-1300 to construct the rice endosperm-specific expression vector pCAMBIA-1300-pMP3-HPV45-L1 and pCAMBIA-1300-pMP3-HPV58-L1.Subsequently,the transgenic rice plants containing HPV-L1 ORF gene were obtained by agrobacterium-mediated genetic transformation of rice callus.The PCR test showed that the HPV45-L1 and HPV58-L1 genes had integrated into the rice genome,suggesting the successful construction of rice endosperm specific expression vectors.

Key words: Human papilloma virus, Transgenic rice, The rice specific endosperm-promoter, Agrobacterium tumefaciens

中图分类号:

Q78
S511.03

王聚财, 刘运超, 陈玉梅, 孟凤丽, 王爱萍, 张二芹, 许倩茹, 邓瑞广, 张改平. HPV45-L1和HPV58-L1蛋白植物表达载体的构建及水稻转基因植株的鉴定[J]. 华北农学报, 2017, 32(2): 55-60. doi: 10.7668/hbnxb.2017.02.009.

WANG Jucai, LIU Yunchao, CHEN Yumei, MENG Fengli, WANG Aiping, ZHANG Erqin, XU Qianru, DENG Ruiguang, ZHANG Gaiping. Construction of HPV45-L1 and HPV58-L1 Plant Expression Vectors and Identification of Transgenic Rice Plants[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(2): 55-60. doi: 10.7668/hbnxb.2017.02.009.

http://www.hbnxb.net/CN/Y2017/V32/I2/55

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HPV45-L1和HPV58-L1蛋白植物表达载体的构建及水稻转基因植株的鉴定

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HPV45-L1和HPV58-L1蛋白植物表达载体的构建及水稻转基因植株的鉴定

Construction of HPV45-L1 and HPV58-L1 Plant Expression Vectors and Identification of Transgenic Rice Plants

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