Variation Analysis Of Hepatitis C Virus Gene Encoding E2 Glycoprotein
By: Saimoon Theeen (2009-VA-565)
| Dr. Muhammad Imran.
Contributor(s): Dr. Sehrish Firyal | Dr. Muhammad Tayyab.
Material type: 
BookPublisher: 2015Description: 106p.Subject(s): Department of Biology and BiotechnologyDDC classification: 2333-T Dissertation note: Hepatitis C virus (HCV) major cause of liver infections was discovered in 1989. It is positive stranded RNA virus and belongs to Flaviviridae family. Its genome shows high rate of variations due to which, its rate of infection is high. As in Pakistan 3% to 6% population and 170 million people worldwide are affected by it. Due to variations in its genome it is classified into 6 major genotypes and these genotypes are further classified into many subtypes. Size of HCV is about 9500 bps which only encodes single polyprotein. This 3000 to 3300 amino acids polyprotein is processed by cellular and viral proteases to generate 10 polypeptides consisting of 4 structural (Core, E1, E2 and P7) and 6 non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A, NS5B). Whereas E2 is considered the most immunogenic gene from all the genes. It involves in the interaction with the host cell and easily escape from the immune system of host due to the presence of hypervariable regions in E2 gene.
To isolate the E2 gene RNA extraction was done using the kit method and then it was converted to the cDNA which is then followed by the PCR amplification. The amplification products were then purify and sent for the sequencing to CAMB. Then the bioinformatics tools were applied on the results. In which the protein structural analysis and epitope mapping was done. Then the conserved epitopes were predicted using the IEDB conservancy analysis tool. The conserved B-cell epitopes (TElAILPCSFTPMPAL and RGERCDIEDRSEQH) and T-cell epitope (TPMPALSTG) are now considered valuable to produce the antibodies against E2 protein.
For diagnosing HCV genotype 3a, these conserved epitopes may be highly useful and may also help in developing a successful vaccine that can target 3a genotypes.
| Item type | Current location | Collection | Call number | Status | Date due | Barcode | Item holds |
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Thesis
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UVAS Library Thesis Section | Veterinary Science | 2333-T (Browse shelf) | Available | 2333-T |
Hepatitis C virus (HCV) major cause of liver infections was discovered in 1989. It is positive stranded RNA virus and belongs to Flaviviridae family. Its genome shows high rate of variations due to which, its rate of infection is high. As in Pakistan 3% to 6% population and 170 million people worldwide are affected by it. Due to variations in its genome it is classified into 6 major genotypes and these genotypes are further classified into many subtypes. Size of HCV is about 9500 bps which only encodes single polyprotein. This 3000 to 3300 amino acids polyprotein is processed by cellular and viral proteases to generate 10 polypeptides consisting of 4 structural (Core, E1, E2 and P7) and 6 non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A, NS5B). Whereas E2 is considered the most immunogenic gene from all the genes. It involves in the interaction with the host cell and easily escape from the immune system of host due to the presence of hypervariable regions in E2 gene.
To isolate the E2 gene RNA extraction was done using the kit method and then it was converted to the cDNA which is then followed by the PCR amplification. The amplification products were then purify and sent for the sequencing to CAMB. Then the bioinformatics tools were applied on the results. In which the protein structural analysis and epitope mapping was done. Then the conserved epitopes were predicted using the IEDB conservancy analysis tool. The conserved B-cell epitopes (TElAILPCSFTPMPAL and RGERCDIEDRSEQH) and T-cell epitope (TPMPALSTG) are now considered valuable to produce the antibodies against E2 protein.
For diagnosing HCV genotype 3a, these conserved epitopes may be highly useful and may also help in developing a successful vaccine that can target 3a genotypes.
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