Construction of a bait plasmid containing HBV PreS1 gene in a yeast two-hybrid system and evaluation of its toxicity and self-activation.
- Author:
Xi ZHANG
1
;
Shu-mei LIN
;
Lie-xiu WU
;
Tian-yan CHEN
;
Feng YE
;
Ying-ren ZHAO
;
Shu-lin ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: Cloning, Molecular; Genetic Vectors; genetics; Hepatitis B Surface Antigens; biosynthesis; genetics; Humans; Plasmids; genetics; Protein Precursors; biosynthesis; genetics; Receptors, Virus; metabolism; Recombinant Fusion Proteins; biosynthesis; genetics; Two-Hybrid System Techniques; Yeasts; genetics; metabolism
- From: Journal of Southern Medical University 2009;29(10):1955-1959
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a yeast expression vector of hepatitis B virus (HBV) PreS1 gene using the Sos-recruitment system (SRS), and evaluate the effect of the expression product on the growth of the yeast cells and activation of the reporter gene.
METHODSThe coding sequence of HBV preS1 was amplified by PCR and cloned into the yeast expression plasmid pSos. The recombinant bait plasmid pSos- PreS1 was verified by sequencing before transformation into competent yeast cells. The effects of the expression product on the yeast cell growth and activation of the reporter gene were evaluated.
RESULTSThe yeast expression vector of HBV PreS1 gene was constructed successfully. The recombinant bait plasmid showed no toxic effect on yeast cdc25H cells without a self-activation of the reporter gene.
CONCLUSIONThe SRS can be used to study the proteins interacting with HBV PreS1 protein and provides a means for obtaining insight into the pathogenic mechanism of HBV.
