HCV vīrusa genomu CORE/ARFP reģiona sekvenču analīze
Date
2015
Authors
Akmeņkalns, Gatis
Journal Title
Journal ISSN
Volume Title
Publisher
Latvijas Universitāte
Abstract
C hepatīta vīruss (HCV) – pieder flavivīrusu dzimtai. HCV ir apvalku saturošs vīruss. HCV var izraisīt tikai cilvēka un augstāko primātu slimības.
C hepatīta vīrusa (HCV) ierosināta infekcija ir pasaulē izplatīts iemesls hroniskām aknu slimībām. Pret HCV vēl nav izstrādāta vakcīna.
HCV ir pirmais vīruss, kas ir atklāts ar molekulārās klonēšanas palīdzību, bez tiešas bioloģisko vai biofizikālo metožu palīdzības.
Galvenā HCV vīrusa īpatnība ir izteikta ģenētiskā mainība.
ARFP (alternatīvās nolasīšanas rāmja proteīns) jeb F (nobīdes), jeb core +1 proteīns (17 kDa) ir radies ribosomu -2/+1 nobīdes rezultātā N-gala core -kodēšanas reģionā HCV poliproteīnā.
Joprojām nav zināma ARFP bioloģiskā funkcija.
Darba mērķis ir apgūt molekulārās bioloģijas metodes.
Noteikt optimālo annealing temperatūru PCR reakcijai core/ARFP reģionam atbilstošo HCV genoma fragmentu amplificēšanai.
Veikt HCV core/ARFP reģiona amplifikāciju un sekvenešanu no HCV hronisko pacientu seruma.
Veikt iegūto PCR produktu klonēšanu un sekvenēšanu.
Analizēt iegūtās sekvences.
Noteikt a-ARFP un a- core antivielu klātbūtni pacientu asins serumos.
Hepatitis C virus (HCV) is member of the Flaviviridae family. HCV is an enveloped virus. HCV can only cause deseases to higher primates and humans. Hepatitis C virus (HCV) infection is a leading cause of chronic liver disease worldwide. Vaccine to HCV is not jet available. HCV was the first virus discovered by molecular cloning without the direct use of biologic or biophysical methods. The main feature of HCV is genetic variability. ARFP alternative reading frame protein), F (frameshift), or core+1 (17 kDa) is the result of ribosomal -2 / +1 shift in N-terminal coding region in the core-HCV polyprotein. The biological function of ARFP in HCV lifecycle is still unknown. The aim of this work is to learn the methods of molecular biology. To determine the optimal temperature of annealing process of PCR reaction core/ARFP specific region of HCV genome fragment amplification. To manage HCV core/ARFP region amplification and sequencing of chronic HCV patient serum. To manage cloning and sequencing of PCR products. To analyze clone sequences. To determin a-ARFP and a- core antibodies in sera prom HCV pacients blood.
Hepatitis C virus (HCV) is member of the Flaviviridae family. HCV is an enveloped virus. HCV can only cause deseases to higher primates and humans. Hepatitis C virus (HCV) infection is a leading cause of chronic liver disease worldwide. Vaccine to HCV is not jet available. HCV was the first virus discovered by molecular cloning without the direct use of biologic or biophysical methods. The main feature of HCV is genetic variability. ARFP alternative reading frame protein), F (frameshift), or core+1 (17 kDa) is the result of ribosomal -2 / +1 shift in N-terminal coding region in the core-HCV polyprotein. The biological function of ARFP in HCV lifecycle is still unknown. The aim of this work is to learn the methods of molecular biology. To determine the optimal temperature of annealing process of PCR reaction core/ARFP specific region of HCV genome fragment amplification. To manage HCV core/ARFP region amplification and sequencing of chronic HCV patient serum. To manage cloning and sequencing of PCR products. To analyze clone sequences. To determin a-ARFP and a- core antibodies in sera prom HCV pacients blood.
Description
Keywords
Bioloģija , hepatīta (HCV) vīruss , genotipi , PCR , ARFP