Construction and expression of six deletion mutants of human astrovirus C-terminal nsP1a/4 protein.
- Author:
Wei ZHAO
1
;
Ke NIU
2
;
Jian ZHAO
3
;
Yi-ming JIN
3
;
Ting-ting SUI
3
;
Wen WANG
3
Author Information
1. Department of Experimental Animal, Institute of Basic Medicine, Liao Ning Medical University, Jinzhou 121200, China. zhaowei8072@163.com
2. First Affiliated Hospital of LiaoNing Medical University, Jinzhou 121200, China.
3. Department of Experimental Animal, Institute of Basic Medicine, Liao Ning Medical University, Jinzhou 121200, China.
- Publication Type:Journal Article
- MeSH:
Amino Acid Motifs;
Astroviridae Infections;
virology;
Humans;
Mamastrovirus;
genetics;
metabolism;
Mutation;
Sequence Deletion;
Transfection;
Viral Nonstructural Proteins;
chemistry;
genetics;
metabolism
- From:
Chinese Journal of Virology
2013;29(5):548-554
- CountryChina
- Language:Chinese
-
Abstract:
Human astrovirus (HAstV) is one of the leading causes of actue virual diarrhea in infants. HAstV-induced epithdlial cell apoptosis plays an important role in the pathogenesis of HAstV infection. Our previous study indicated that HAstV non-structural protein nsPla C-terminal protein nsPla/4 was the major apoptosis functional protein and probably contained the main apoptosis domains. In order to screen for astrovirus encoded apoptotic protien, nsPla/4 and six turncated proteins, which possessed nsPla/4 protein different function domain ,were cloned into green fluorescent protein (GFP) vector pEG-FP-N3. After 24-72 h transfection, the fusion protein expression in BHK21 cells, was analysis by fluorescence microscope and Western blot. The results indicated seven fusion proteins were observed successfully in BHK21 cell after transfected for 24 h. Western blot analysis showed that the level of fusion protein expressed in BHK21 cells was increased significantly at 72h compared to 48h in transfected cells. The successful expression of deletion mutants of nsPla/4 protein was an important foundation to gain further insights into the function of apoptosis domains of nsPla/4 protein and it would also provide research platform to further confirm the molecule pathogenic mechanism of human astrovirus.
