[Identification of the interactions between JTV1 and NLS-RAR alpha in vivo and in vitro].
OBJECTIVE
To identify the interactions between JTV1 and NLS-RAR alpha by Yeast two-hybrid and co-immunoprecipitation.


METHODS
The plasmids of bait-protein and JTV1 protein were cotransformed into yeast AH109 to investigate their interactions in vivo. Tagged fusion protein eukaryotic expression vectors were constructed and then cotransfected into human embryo kidney 293 cells. Co-immunoprecipitation was used to investigate the interactions between NLS-RAR alpha and JTV1 in vitro.


RESULTS
Blue clones were found in QDO/X-alpha-gal plates. Eukaryotic expression vectors were co-transfected into HEK 293 cells. The HA-NLS-RAR alpha protein was immunoprecipitated by anti-HA polyclonal antibody. Myc-JTV1 protein was detected by western blotting with anti-Myc monoclonal antibody from the immunoprecipitared complex.


CONCLUSIONS
The interactions between NLS-RAR alpha and JTV1 are identified by Yeast two-hybrid and co-immunoprecipitation.
Wang
Chong
C
Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Faculty of Laboratory Medicine in Chongqing Medical University, Chongqing 400016, China.
Wang
Dong-sheng
DS
Liu
Bei-zhong
BZ
Hao
Po
P
Liu
Chang
C
Jin
Dan-ting
DT
Zhong
Liang
L
Wang
Chun-guang
CG
D045744:Cell Line, Tumor; D006801:Humans; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D025941:Protein Interaction Mapping; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D000072482:Retinoic Acid Receptor alpha; D014162:Transfection; D020798:Two-Hybrid System Techniques
D009363:Neoplasm Proteins; C000607071:RARA protein, human; D018168:Receptors, Retinoic Acid; D011993:Recombinant Fusion Proteins; D000072482:Retinoic Acid Receptor alpha