Localization of functional domains of HEV ORF1 in cells.
- Author:
Hui HUANG
1
;
Zi-Zheng ZHENG
;
Min ZHAO
;
Jing-Xian LI
;
Wang-Sheng LAI
;
Ji MIAO
;
Jun ZHANG
;
Ning-Shao XIA
Author Information
1. National Institute of Diagnostics and Vaccine Development in Infectious Disease, School of Life Sciences, Xiamen University, Xiamen 361005, China. huanghui@xmu.edu.cn
- Publication Type:Journal Article
- MeSH:
Blotting, Western;
Cells, Cultured;
Hepatitis E virus;
genetics;
Humans;
Open Reading Frames;
Viral Proteins;
genetics;
physiology
- From:
Chinese Journal of Virology
2011;27(3):195-201
- CountryChina
- Language:Chinese
-
Abstract:
To investigate the expression and localization of various functional domains of ORF1 polyprotein and ORF3 protein of hepatitis E virus in host cells, the coding sequences of the various functional domains (RdRp, HEL, MET, PLP, X) of ORF1 were separately cloned into pcDNA3. 1-GFP vectors for constructing the recombinant plasmids which were verified by enzyme digestion and sequencing. The exact expression of the fusion proteins were detected by Western Blot, and the distribution and localization were observed by the laser scanning confocal microscope(LSCM). In huh7 cells, GFP-RdRp proteins were found mainly in the nuclei, GFP-HEL proteins were distributed vesicularly around the nucleus, GFP-MET proteins were distributed granularly both in the nuclei and the cytoplasm, GFP-PLP proteins had polar distribution around the nucleus, and unknown GFP-X proteins were distributed uniformly both in the nuclei and the cytoplasm. Different localization of these proteins verified the previous data obtained from in vitro studies, providing a support for further research on the biological functions of various proteins coded by HEV genome.
