Screening and cloning of hepatitis C virus non-structural protein 4B interacting protein gene in hepatocytes / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
; (6): 248-251, 2005.
Artigo
em Chinês
| WPRIM (Pacífico Ocidental)
| ID: wpr-333030
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To investigate biological functions of non-structural protein 4B (NS4B) of hepatitis C virus (HCV), yeast-two hybrid technique was performed to seek proteins in hepatocytes interacting with HCV NS4B.</p><p><b>METHODS</b>HCV NS4B bait plasmid was constructed by ligating the NS4B gene with carrier plasmid pGBKT7 and transformed into yeast cells AH109 (type alpha). The transformed yeast cells were amplified and mated with yeast cells Y187 (alpha type) containing liver cDNA library plasmid pACT2 in 2 x YPDA medium. Diploid yeast cells were 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. After extracting plasmid from blue colonies, plasmid DNA was transformed into competent Escherichia coli and analysed by DNA sequencing and bioinformatics.</p><p><b>RESULTS</b>Five genes in eight positive colonies were obtained. There were one NADH dehydrogenase subunit 3, one cytochrome c oxidase subunit III, one retinol binding protein 4, one reticulon 3-A (RTN3) and one fibrinogen gamma polypeptide (FGG).</p><p><b>CONCLUSION</b>Genes of HCV NS4B interacting proteins in hepatocytes were successfully cloned and the results paved the way for studying the biological functions of NS4B and associated proteins.</p>
Texto completo:
Disponível
Contexto em Saúde:
ODS3 - Saúde e Bem-Estar
Problema de saúde:
Meta 3.3: Acabar com as doenças tropicais negligenciadas e combater as doenças transmissíveis
Base de dados:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Ligação Proteica
/
Proteínas de Ligação ao Retinol
/
Proteínas de Transporte
/
Biblioteca Gênica
/
Western Blotting
/
Clonagem Molecular
/
Proteínas não Estruturais Virais
/
Complexo IV da Cadeia de Transporte de Elétrons
/
Técnicas do Sistema de Duplo-Híbrido
Tipo de estudo:
Estudo diagnóstico
/
Estudo de rastreamento
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Experimental and Clinical Virology
Ano de publicação:
2005
Tipo de documento:
Artigo