JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2015, Vol. 53 ›› Issue (9): 35-40.doi: 10.6040/j.issn.1671-7554.0.2015.096

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Construction of the minireplicon of enterovirus 71

MA Yingwei, SUN Lele, LI Jing, QIAO Qiao, ZHUANG Zhichao, ZHAO Li, YU Xuejie, WANG Zhiyu, WEN Hongling, SUN Yanlin   

  1. Department of Virology, School of Public Health, Shandong University, Jinan 250012, Shandong, China
  • Received:2015-01-27 Online:2015-09-10 Published:2015-09-10
  • Contact: 温红玲。E-mail:wenhongling@sdu.edu.cn E-mail:wenhongling@sdu.edu.cn

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Abstract: Objective To construct the minireplicon of non-structural protein and 3C protease of EV71 and study the replication and expression of the minireplicon. Methods The genes of the enhanced green fluorescent protein(EGFP), amplified by PCR, and the genes of non-structural protein and 3C protease were inserted into pMD19-T vector through specific restriction sites and ligase. The expression and replication of the minireplicon were detected by fluorescence microscope and checked by real-time PCR respectively. Results Specific green fluorescence was detected after in vitro transcription and transfection of the plasmid containing the genes of EV71 non-structural proteins and the 3C protease, and there were significant differences in the intensity of the fluorescence among different groups. The content of the RNA remained same between 12 h to 72 h after transfection. Conclusion The abilities of transcriptions of the full-length non-structural protein and the 3C protease were different. The replication of the minireplicon could not be detected in RD cells, which suggested the importance of the structural protein to the replication of the virus.

Key words: Minireplicon, Non-structural protein, Enterovirus 71, Reporter gene

CLC Number: 

  • R373.2
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