Screening and identification of proteins interacting with RAR alpha-V via yeast two-hybrid system.
- Author:
Dong-sheng WANG
1
;
Chong WANG
;
Bei-zhong LIU
;
Qian-feng XIA
;
Po HAO
;
Chang LIU
;
Dan-ting JIN
;
Liang ZHONG
Author Information
- Publication Type:Journal Article
- MeSH: Gene Library; Humans; K562 Cells; Protein Interaction Mapping; Receptors, Retinoic Acid; metabolism; Retinoic Acid Receptor alpha; Two-Hybrid System Techniques
- From: Chinese Journal of Hematology 2008;29(6):388-392
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo screen the protein interacting with retinoic acid receptor variant protein (RAR alpha-V) via the yeast two-hybrid technique (YTHT) and to find out the targets protein and study its biological function.
METHODSThe bait vector of pGBKT7-RAR alpha-V was constructed for screening proteins interacting with RAR alpha-V in K562 cell cDNA expression library via YTHT. The protein-protein interaction was confirmed with re-transformation in yeast and GST pull-down in vitro.
RESULTSThe bait vector was successfully constructed without toxicity, leakage and self-activation. Sixteen proteins were screened by YTHT and eight positive clones were identified by re-transformation in yeast. The interaction between RAR alpha-V and JTV-1 was confirmed by GST pull-down in vitro.
CONCLUSIONSThere are some kinds of proteins interacting with RAR alpha-V in cell. The biological dysfunction caused by certain protein-protein interaction may be involved in the pathogenesis of leukemia.
