Gene Clone, Expression And Study On Immunogenity And Mimotope Of Hepatitis C Virus Envelope Protein E2

Hepatitis C virus (HCV) is the major etiologic agent of posttransfusion and community-acquired non-A, non-B hepatitis. Gene encoding HCV envelope protein E2 is high variable, which can cause an immune escape of the virus and is related with chronic HCV infection. E2 protein, including hypervariable region 1 (HVR1) in the N-terminal region, contains some epitopes that can induce neutralizing antibodies. The research on gene structure and protein function of E2 region will be helpful for prognosis, prevention and treatment of HCV infection.In this study, gene of envelope protein E2 was isolated from one HCV-infected person. Then it was cloned and expressed in procaryotic and eukaryotic expression systems. The purified E2 protein was used to study its immunogenity and mimotope.1. Cloning and Sequencing of HCV Envelope Protein E2HCV RNA was extracted from the serum of one HCV-infected blood donor from Hebei Province, China. The envelope protein E2 gene was amplified by RT-PCR and cloned into expressing plasmid pQE30 and pTrxFus. By induction of E2 protein, two clones were selected and sequenced. The results showed that the genes were envelope protein E2 genes of HCV genotype 1b.2. Expression and Purification of HCV E2 ProteinA series of expressing plasmids were constructed in procaryotic and eukaryotic expression systems. In pQE30 expression system, three E2 proteins were expressed and the molecular weight was 16kD, 30kD and 38kD respectively. The protein was purified with Nickel chelate affinity chromatography. The purified protein can react specifically with sera from HCV-infected patients by ELISA. In eukaryotic expression system, some of the constructed plasmids could express E2 proteins, but most of them couldn't express the desired proteins.3. Study on Immunogenity and Mimotope of HCV Envelpoe Protein E2...