ABSTRACT
<p><b>BACKGROUND</b>To express recombinant non-structural protein 3 of hepatitis C virus (HCV) in E. coli.</p><p><b>METHODS</b>The non-structural 3 (NS3) region DNA fragment of HCV was amplified by polymerase chain reaction (PCR) and inserted into inducible proeukaryotic expressive vector pET 30C(+)at Bam H1/EcoR1 sites. The competent BL21 (DE3) E.coli was transformed, and then cultured and induced with IPTG. The expressed HCV NS3 protein was confirmed with ELISA and dot blot hybridization using HCV NS3-specific single chain Fv (ScFv) antibody.</p><p><b>RESULTS</b>1 893 bp DNA fragment of HCV NS3 coding region was amplified by PCR technique. HCV NS3 expressive vector pET-NS3 was constructed. After transformation with pET-NS3 and induction with IPTG, recombinant HCV NS3 protein was expressed and confirmed by specific ELISA and dot blot hybridization.</p><p><b>CONCLUSIONS</b>The recombinant HCV NS3 can be expressed in E. coli.</p>
Subject(s)
Escherichia coli , Genetics , Gene Expression , Hepatitis C Antibodies , Genetics , Polymerase Chain Reaction , Recombinant Proteins , Genetics , Viral Nonstructural Proteins , GeneticsABSTRACT
In order to develop ELISA method for measuring the degradation fragment of fibronectin (MAD2),hybridoma technique was used to obtain the monoclonal antibody (McAb)IgG 1 against MAD2 without cross reaction with fibronectin.The sera from 277 patients with hepatocellular carcinoma (HCC),76 with metastatic hepatic carcinoma (HMC),98 with alimentary canal carcinoma (ACC) and 156 with chronic liver disease(CLD) and 48 healthy subjects were assayed with ELISA method using this antibody.The examination showed that the mean value of MAD2 from the patients with HCC showed obviously significant difference compared with those of CLD,HMC,ACC and normal control groups ( p
ABSTRACT
Pre-S2 protein in 78 samples of liver from patients with various types of hepatitis B was determined by immunohistochemical method.45 out of 78 samples (57.69%) were positive for pre-S2.The location and expression patterns of pre-S2 were similar to those of HBsAg,i.e.,pre-S2 was found in cytoplasm of hepatocytes,epithelial cells of bile ducts,and hepatocellular carcinoma cells,etc,Four different patterns diffuse,inclusion,submembranous,and membranous type were demonstrated.The membranous expression of pre-S2 was often associated with activity of liver diseases.The positive rate of pre-S2 was significantly higher in HBcAg positive group than HBcAg negative group.It seems that pre-S2 protein of HBV may be a replicative marker of HBV.