Verification of interaction between glutamate-ammonia ligase and nuclear localization signal-retinoic acid receptor α protein inside and outside cells
10.3724/SP.J.1008.2010.00468
- Author:
Yan WU
1
Author Information
1. Ministry of Education
- Publication Type:Journal Article
- Keywords:
Co-immunoprecipitation;
Glutamate-ammonia ligase;
Nuclear localization signals;
Protein interaction;
Retinoic acid receptor α;
Two-hybrid system techniques
- From:
Academic Journal of Second Military Medical University
2010;31(5):468-471
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To verify the interaction between glutamate-ammonia ligase (GLUL) and nuclear localization signal-retinoic acid receptor α (NLS-RARα) protein by yeast two-hybrid and co-immunoprecipitation method. Methods: The two plasmids expressing NLS-RARα bait-protein and GLUL protein were co-transformed into yeast AH109 to investigate the interaction in vivo. Tagged fusion protein eukaryotic expression vectors were constructed and co-transfected into HEK 293 cells. Co-immunoprecipitation was used to investigate the interaction between NLS-RARα and GLUL in vitro. Results: Positive blue clones were found in the QDO/X-α-gal plate. Eukaryotic expression vectors were co-transfected into HEK 293 cells, then HA-NLS-RARα protein was immunoprecipitated by anti-HA polyclonal antibody, and GLUL-cMyc protein expression was confirmed by Western blotting analysis using anti c-Myc monoclonal antibody. Conclusion: The interaction between NLS-RARα and GLUL has been verified by both yeast two-hybrid and co-immunoprecipitation.
