Dissertation > Excellent graduate degree dissertation topics show

Inhibition of HCV Translation by HCV NS3Helicase RNA

Author: LiuXiaoLei
Tutor: LiuShuiPing
School: Central South University
Course: Medical Microbiology
Keywords: HCV NS3helicase mutation plasmid luciferase translationinitiation
CLC: R512.63
Type: Master's thesis
Year: 2013
Downloads: 2
Quote: 0
Read: Download Dissertation

Abstract


Hepatitis C virus (HCV) is a major cause of chronic hepatitis and the leading cause of end-stage liver cirrhosis and hepatocellular carcinoma. The best available HCV antiviral therapy is a combination of pegylated interferon-α(IFN-α) and ribavirin-based therapy.However, because of its side effect, high cost and low sustained virological response, searching for a more effective anti-HCV therapeutic method is considered to be urgent. Due to the low level of HCV replication in vivo, it is hard to build a satisfactory and efficient replication and infection model in vitro. The study on HCV life cycle and regulation of internal gene expression is becoming increasingly significant. This research aims to study the influence of HCV IRES translation initiation activity by HCV NS3helicase as well as discussing the self-regulation mechanism of HCV translation.Methods:(1) HCV NS3full sequence, HCV NS3helicase, its amino acid mutation and defective expression plasmids were constructed;(2) HCV IRES-driven luciferase expression plasmid was transfected to293T cell model, based on which, constructed plasmids were transfected into these cells by utilizing the calcium phosphate transfection method. Luciferase expressing activity was then detected in24h;(3) The total RNA was extracted, RT-PCR is applied to detect expressions of pCMVNCRLuc, cell IFN-β and PKR mRNA relative expression level.Results:(1) Results of plasmid enzyme digestion and sequencing demonstrate that recombination HCV NS3helicase, its amino acid mutation and defective expression plasmids were constructed;(2) After in vitro transfection, HCV NS3helicase and its amino acid mutation plasmid can inhibit the expression of luciferase in transfected cells; Similarly, HCV NS3helicase defective expression plasmid can inhibit the expression of luciferase in transfected cells;(3) Semi-quantitative RT-PCR detecting of HCV pCMVNCRLuc and pRL-TK mRNA relative expression showed no significant difference. IFN-J3was not detected in cells, indicating HCV NS3helicase typically plays an inhibiting role on HCV translation. The mRNA expression of PKR showed no significant.Conclusions:(1) HCV NS3helicase, its amino acid mutation and defective expression plasmids were constructed successfully;(2) HCV IRES translation initiation activity was inhibited by HCV NS3helicase RNA;(3) HCV IRES translation initiation activity was inhibited by HCV NS3helicase RNA, which can be done not by increasing PKR expression level. Its mechanism needed to be further studied.

Related Dissertations

  1. Promoter Activity Analysis of Cytochrome P450 Gene CYP9A17v2 from Helicoverpa Armigera (H(?)bner),S435.622
  2. Development of ATP Bioluminescent Method and Its Kit for the Detection of Bacterial Count in Food,TS207.4
  3. HCV NS2TP Gene Regulation Mechanism,R512.63
  4. A Research of the Incidence of HIV/HCV/HBV/syphilis and Risk Behaviors among Injecting Drug Users in Xichang County of Sichuan Province,R181.3
  5. Influence of CD8~+ T Cell Pressure on HCV Evolution,R392
  6. The Rapid Toxicity Testing Based on Luciferase and the Study of on-line Toxicity Biosensor,X85
  7. Differential Expression of MS4A7 Gene in U-937 Differentiation and Identification of Its Promoter,R733.71
  8. HCV Viral Load and Subtypes among HCV Mono-infected and HIV/HCV Co-infected Patients,R512.63
  9. Effect of Hepatitis C Virus NS5A Protein on TLR3,TLR4,R512.63
  10. Study on the Cleavage Activity of the Optimized M1GS Ribozymes That Target the Core Gene of Hepatitis C Virus,R512.63
  11. Screen the Differential Expression of Tyrosine-Phosphorylated Proteins Involved in Hepatocyte Transfected by HCV/NS3 Plasmid,R363
  12. Modulation of IFN-γ-induced TRIM22 Expression by Histone Deacetylase Inhibitor TSA and the Underlying Mechanism,R346
  13. Effect of TLR4 Promoter Polymorphisms on Transcription Activity in Neonates,R346
  14. The Biology, Ecology and Molecular Evolution Study of Chetoneura Shennonggongensis,S433
  15. Influence of Hepatitis C Virus Genotypes and F Protein on Chronic Hepatitis C Virus Infection and Hepatocellular Carcinom Cell Apoptosis,R735.7
  16. Influence of DNA Polymerase β Promoter Mutation to Its Transcriptional Activity in Esophageal Carcinoma Cell EC-1,R735.1
  17. Investigation on Blood Supersession in Jinan Area and Research on the Countermeasures,R197.6
  18. Study on the Bacterial Luciferase Bioluminescent System in Vitro and Its Application in Detection of Pathogenic Bacteria in Fishery Products,TS254.7
  19. The Establishment of HCV RNA Detection Methods & the Analysis of Variation Characteristic on Drug Target,TQ460.1
  20. Establishment and Verification of Human Lung Cancer Cell for Application in Vivo Imaging System,R734.2
  21. Preliminary Study of Mutation of DNA polβ Promoter in the Esophageal Carcinoma Tissue,R735.1

CLC: > Medicine, health > Internal Medicine > Infectious disease > Viral infections > Viral Hepatitis > Hepatitis C (non- A non-B )
© 2012 www.DissertationTopic.Net  Mobile