[Screening and identification of proteins interacting with RAR alpha-V via yeast two-hybrid system].
OBJECTIVE
To 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.


METHODS
The 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.


RESULTS
The 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.


CONCLUSIONS
There 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.
Wang
Dong-sheng
DS
Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Faculty of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
Wang
Chong
C
Liu
Bei-zhong
BZ
Xia
Qian-feng
QF
Hao
Po
P
Liu
Chang
C
Jin
Dan-ting
DT
Zhong
Liang
L
D015723:Gene Library; D006801:Humans; D020014:K562 Cells; D025941:Protein Interaction Mapping; D018168:Receptors, Retinoic Acid / Q000378:metabolism*; D000072482:Retinoic Acid Receptor alpha; D020798:Two-Hybrid System Techniques
C000607071:RARA protein, human; D018168:Receptors, Retinoic Acid; D000072482:Retinoic Acid Receptor alpha