肠道病毒71型（EV71）荧光病毒的构建

生物技术通报 ›› 2014, Vol. 0 ›› Issue (7): 179-183.

肠道病毒71型（EV71）荧光病毒的构建

朱守海^{1, 2}, 王璐璐^{1, 2}

1.天津科技大学生物工程学院, 天津 300457；
2.天津市国际生物医药联合研究院, 天津 300457

收稿日期:2013-12-11 出版日期:2014-07-15 发布日期:2014-07-16
作者简介:朱守海, 男, 硕士, 研究方向：生物化学, E-mail：zhusshh@163.com；王璐璐同为本文第一作者

The Construction of the EGFP Enterovirus 71

Zhu Shouhai^{1, 2} Wang Lulu^{1, 2}

1. College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457；
2.Tianjin International Joint Academy of Biomedicine, Tianjin 300457

Received:2013-12-11 Published:2014-07-15 Online:2014-07-16

摘要/Abstract

摘要： 人肠道病毒71型（Enterovirus, EV71）是引起手足口病的主要病原体。首先利用反向遗传学的方法, 构建了EV71全长cDNA感染性克隆, 经体外转录、转染RD细胞后成功获得了拯救病毒。随后, 将增强型绿色荧光蛋白基因（Enhanced green fluorescent protein, EGFP）插入到EV71基因组中构建荧光病毒。结果表明, 此荧光病毒不仅可以侵染、复制, 在传代的过程中EGFP基因也保持了较好的稳定性, 表明其可以作为一种报告病毒应用于高通量的EV71抗病毒药物筛选中。

关键词: EV71, 感染性克隆, 增强型绿色荧光蛋白, 荧光病毒

Abstract: Human Enterovirus 71（EV71）is one of the major agents causing hand, foot and mouth disease（HFMD）. An infectious full-length cDNA clone of EV71 as well as the recuse virus were obtained using the reverse genetic techniques. Then, the EGFP gene was inserted into the EV71 genome to construct the reporter virus. The results show that the EGFP-EV71 virus not only are replication-competent and infectious, but the EGFP gene are stable during passaging, implicating the recombinant virus could be used as a reporter virus for high throughput EV71 antiviral drugs screening.

Key words: EV71, Infectious cDNA clone, EGFP, Reporter virus

朱守海, 王璐璐. 肠道病毒71型（EV71）荧光病毒的构建[J]. 生物技术通报, 2014, 0(7): 179-183.

Zhu Shouhai Wang Lulu. The Construction of the EGFP Enterovirus 71[J]. Biotechnology Bulletin, 2014, 0(7): 179-183.

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