[Identification of interaction between BPHL and PML-C].
Sichuan Da Xue Xue Bao Yi Xue Ban
; 44(1): 6-9, 2013 Jan.
Article
em Zh
| MEDLINE
| ID: mdl-23600199
ABSTRACT
OBJECTIVE:
To explore the interaction between BPHL and PML-C by co-immunoprecipitation and yeast two-hybird system.METHODS:
The recombination expression plasmids pGBKT7-PML-C and pACT2-BPHL were cotransformed into yeast AH109, to investigate their interaction in vivo. The expression vector of HA-tagged fusion protein (pCMV-HA-PML-C) and the expression vector of myc-tagged fusion protein (pCMV-myc-BPHL) were constructed and identified respectively, and cotransfected into human embryo kidney 293 (HEK293) cells. The interaction between PML-C and BPHL was investigated by co-immunoprecipitation in vitro.RESULTS:
Blue clones were found in QDO/5-bromo-4-chloro-3-indolyl-alpha-D-galactoside (X-alpha-gal) plate, eukaryotic expression vectors named as pCMV-HA-PML-C and pCMV-myc-BPHL were constructed and confirmed with double restriction enzyme digestion and co-transfected into HEK 293 cells successfully. After immunoprecipitation of HA-PML-C with anti-HA polyclonal antibody, expressed myc-BPHL protein was identified by Western blot with anti-c-myc monoclonal antibody from immunoprecipitated complex.CONCLUSION:
The eukaryotic expression vector of PCMV-HA-PML-C and PCMV-myc-BPHL were constructed successfully. The interaction between PML-C and BPHL was identified by co-immunoprecipitation and yeast two-hybird technique.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Fatores de Transcrição
/
Proteínas Nucleares
/
Hidrolases de Éster Carboxílico
/
Proteínas Supressoras de Tumor
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Humans
Idioma:
Zh
Revista:
Sichuan Da Xue Xue Bao Yi Xue Ban
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
China