Isolation of Tara protein and its gene cloning / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
; (6): 486-490, 2004.
Article
en Zh
| WPRIM
| ID: wpr-353276
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To isolate and identify TRF1 immunoprecipitating protein complex and to clone the candidate gene.</p><p><b>METHODS</b>The co-immunoprecipitation assay was employed to isolate TRF1 protein complex and the immunoprecipitate was subjected to MALDI-TOF mass spectrometry for protein identification. The candidate gene was amplified by temperature-gradient PCR from human testis cDNA library and then cloned into pEGFP-C2 vector for eukaryotic expression. The amplified gene was verified by direct sequencing and GFP-tagged protein was confirmed by immunoblotting.</p><p><b>RESULTS</b>Tara protein with the size of 68 kD was identified from the TRF1 precipitate. The candidate gene amplified from cDNA library was about 1.7 kb as expected. Sequencing demonstrated the amplified fragment had 99.9% of homogenesis with Tara CDS sequence (gi:30474869). GFP-tagged fusion protein was about 100 kD. Tara was diffusely distributed in cytoplasm at interphase and in whole cells at mitotic phase.</p><p><b>CONCLUSION</b>Tara might be an interacting protein with TRF1. However, further investigation would be required to confirm if they were bona fide partners.</p>
Texto completo:
1
Índice:
WPRIM
Asunto principal:
Unión Proteica
/
Células HeLa
/
Química
/
Clonación Molecular
/
Proteína 1 de Unión a Repeticiones Teloméricas
/
Genética
/
Metabolismo
/
Proteínas de Microfilamentos
Tipo de estudio:
Prognostic_studies
Límite:
Humans
Idioma:
Zh
Revista:
Journal of Zhejiang University. Medical sciences
Año:
2004
Tipo del documento:
Article