Screening and identification of proteins interacting with HCV NS4A via yeast double hybridization in leukocytes and gene cloning of the interacting protein / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 47-49, 2007.
Artículo
en Chino
| WPRIM
| ID: wpr-305499
ABSTRACT
<p><b>OBJECTIVE</b>To screen proteins interacting with HCV NS4A protein in leukocytes by yeast-double hybridization.</p><p><b>METHODS</b>The bait plasmid pGBKT7-NS4A was transformed into yeast AH109 was transformed, and the expressing of the fusion protein was identified by SDS-page. The transformed yeast was mated with yeast Y187 containing leukocytes cDNA library plasmid in 2xYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selecting two times and screening. After extracting and sequencing of plasmid from blue colonies, analysis was conducted by bioinformatics. And, the gene encoding the interesting protein was cloned, and back-cross was performed.</p><p><b>RESULTS</b>Forty-five colonies were sequenced, among them, 29 colonies were human calcium modulating cyclophilin ligand (CAML). The gene encoding CAML was cloned, and the interaction between NS4A and CAML was ensured.</p><p><b>CONCLUSION</b>Seven kinds of proteins interacting with NS4A in leukocytes were successfully screened and the results brought some new clues for studying the pathogenesis of HCV.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Unión Proteica
/
Transformación Genética
/
Proteínas Virales
/
Proteínas Portadoras
/
Biblioteca de Genes
/
Clonación Molecular
/
Proteínas no Estructurales Virales
/
Técnicas del Sistema de Dos Híbridos
/
Biología Celular
/
Proteínas Adaptadoras Transductoras de Señales
Tipo de estudio:
Estudio diagnóstico
/
Estudio de tamizaje
Límite:
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Experimental and Clinical Virology
Año:
2007
Tipo del documento:
Artículo
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