A study of the down-regulation effect of hepatitis B virus pre-protein S2 on inducible nitric oxide synthase gene promoter / 中华肝脏病杂志
Chinese Journal of Hepatology
;
(12): 749-753, 2005.
Artigo
em Chinês
| WPRIM
| ID: wpr-276363
ABSTRACT
<p><b>OBJECTIVES</b>To investigate the regulating effect of HBV pre-S2 protein on iNOS gene promoter and the molecular biological mechanisms of pre-S2 protein in HBV pathogenicity.</p><p><b>METHODS</b>Polymerase chain reaction (PCR) technique was employed to amplify the sequence of iNOS promoter and 3 deletion mutants using HepG2 genomic DNA as the template, and the products were cloned into the pGEM-T vector. The iNOS gene and 3 deletion mutants were cut from T- iNOS by Kpn I and Xho I, and then cloned into pCAT3-Basic. The resulting vectors were named p1-iNOSp, p2-iNOSp, p3-iNOSp, and p4-iNOSp. Each of the reporter vectors was transfected into the HepG2 cell line and cotransfected into HepG2 cells with pcDNA3.1(-)-pre-S2 by FuGENE 6 transfection reagents. The HepG2 cells transfected with pCAT3-Basic were used as a negative control. The activity of CAT in HepG2 cells transfected was detected by an ELISA kit 48 hours after the transfection, which reflected the regulating effect of HBV pre-S2 protein on iNOS gene promoter activity.</p><p><b>RESULTS</b>The expressive vector pcDNA3.1(-)-pre-S2 and report vector pCAT3-iNOSp were constructed and confirmed by restriction enzyme digestion and sequencing. The expression of pcDNA3.1(-)-pre-S2 in HepG2 cells could down-regulate the activity of p1-iNOSp, p3-iNOSp, and the inhibition rate was 54.7% and 79.5%, respectively. The expression of pcDNA3.1(-)-pre-S2 in HepG2 cells had no regulatory effects on p2-iNOSp and p4-iNOSp.</p><p><b>CONCLUSION</b>It is suggested that HBV pre-S2 protein can down-regulate iNOS gene promoter.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Precursores de Proteínas
/
Transfecção
/
Regulação para Baixo
/
Ativação Transcricional
/
Regiões Promotoras Genéticas
/
Óxido Nítrico Sintase Tipo II
/
Genética
/
Antígenos de Superfície da Hepatite B
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Hepatology
Ano de publicação:
2005
Tipo de documento:
Artigo
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