Screening of hepatocyte proteins interacting with hepatitis B virus X protein using CytoTrap yeast two-hybrid technique.
- Author:
Baihai JIAO
1
;
Yan WEN
;
Xiaojia LIU
;
Yue FENG
;
Amei ZHANG
;
Li LIU
;
Xueshan XIA
Author Information
- Publication Type:Journal Article
- MeSH: Genetic Vectors; Hepatocytes; metabolism; Humans; Plasmids; Protein Interaction Domains and Motifs; Proteins; metabolism; Trans-Activators; metabolism; Two-Hybrid System Techniques
- From: Journal of Southern Medical University 2013;33(4):486-490
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo screen the hepatocyte proteins that interact with hepatitis B virus X protein (HBx).
METHODSThe recombinant plasmid pSos-HBx was constructed by inserting Sos-HBx fragment into the bait vector, and after sequence verification the plasmid was transformed into competent yeast cells. The expression and self-activation of Sos-HBx protein was detected in the yeast cells. The hepatocyte proteins interacting with the bait protein was screened with CytoTrap yeast two-hybrid technique.
RESULTSThe reconstructed plasmid harboring HBx gene expressed Sos-HBx protein in the yeast cells without self-activation of the protein. CytoTrap yeast two-hybrid system identified 6 hepatocyte proteins that interacted with HBx, including fibronectin 1, translationally controlled tumor protein, IQ motif and WD repeats 1, follistatin, orosomucoid 1, and disulfide isomerase family A member 3.
CONCLUSIONSix HBx-binding hepatocyte proteins have been identified using the CytoTrap yeast two-hybrid system, which provides clues for further investigation of the role of HBx protein in hepatitis and liver cancer.
