Up-regulating effect of hepatitis C virus core protein on inducible nitric oxide synthase gene promoter
- VernacularTitle:丙型肝炎病毒核心蛋白上调诱导型一氧化氮合酶基因启动子的转录活性
- Author:
Feng-Jin GUO
;
Jun CHENG
;
Yan LIU
;
- Publication Type:Journal Article
- Keywords:
Hepacivirus;
Promoter regions(genetics);
Nitric-oxide synthase;
Viral coreproteins;
Transerption,genetic
- From:
Chinese Journal of Infectious Diseases
1997;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the transactivating effect of hepatitis C virus(HCV)core protein on inducible nitric oxide synthase(iNOS)gene promoter and the molecular biological mecha- nisms of HCV pathogenesis.Methods Polymerase chain reaction(PCR)technique was employed to amplify the sequence of iNOS promoter by using HepG2 genomic DNA as template,and the product was cloned into pGEM-T vector.The iNOSp gene was cut from T-iNOSp by KpnⅠand XhoⅠ,and then was cloned into pCAT3-Basic,the constructed vector was named as pCAT3-iNOSp,pCAT3-iN- OSp was transfected into the LO_2 cell line.LO_2 cell was also cotransfected with pcDNA3.1(-)-core and pCAT3-iNOSp by FuGENE 6 transfection reagents.The LO_2 cells transfected with pCAT3-Basic was used as negative control.The activity of CAT in LO_2 cells was detected by an ELISA kit after 48 hours,which reflected the transactivating function of HCV core protein to iNOS gene promoter.Re- sults The expressive vector pcDNA3.1(-)-core and report vector pCAT3-iNOSp had been construc- ted and confirmed by restriction enzyme digestion and sequencing.The expression of CAT in LO_2 cells transfected with pCAT3-iNOSp and peDNA3,1(-)-core was 11 times as higher as that of pCAT3-bas- ic,and 6 times as higher as that of pCAT3-iNOSp.Conclusion It is suggested that HCV core protein can transactivate iNOS gene promoter.
