Cover Image

Generation of gene knockout cassette (GKO-AbR) for claudin-1 gene targeting / Laily Norsafira Sahizan

Hepatitis C is a type of inflammation of the liver, origin of Hepatitis C Virus (HCV) and it is estimated that more than 170 million people worldwide have been infected with this virus. Claudin-I (CLDN-1) is one of the receptors that are believed to be crucial for HCV entry process, but the details...

Full description

Saved in:
Bibliographic Details
Main Author: Sahizan, Laily Norsafira
Format: Thesis
Language:English
Published: 2013
Subjects:
Pharmacopoeias
Online Access:https://ir.uitm.edu.my/id/eprint/109158/1/109158.PDF
https://ir.uitm.edu.my/id/eprint/109158/
Tags: Add Tag
No Tags, Be the first to tag this record!
id my.uitm.ir.109158
record_format eprints
spelling my.uitm.ir.1091582025-02-21T23:45:38Z https://ir.uitm.edu.my/id/eprint/109158/ Generation of gene knockout cassette (GKO-AbR) for claudin-1 gene targeting / Laily Norsafira Sahizan Sahizan, Laily Norsafira Pharmacopoeias Hepatitis C is a type of inflammation of the liver, origin of Hepatitis C Virus (HCV) and it is estimated that more than 170 million people worldwide have been infected with this virus. Claudin-I (CLDN-1) is one of the receptors that are believed to be crucial for HCV entry process, but the details on the mechanisms are still unclear. This research aims to design the primers and amplify the antibiotic resistance gene of interest from the commercially available plasmid and to produce gene knockout cassette of claudin-I by linking the homology arms of exon 1 claudin-I region. PrimerBlast online software was used as the tool to design the target-specific primers ofpcDNA3.l and pCMV/Bsd vector. The gene of interest used from the pcDNA3.1 vector was Neomycin resistance gene while from pCMV/Bsd vector the gene of interest used was Blasticidin resistance gene. The sizes of the primers produced for pcDN A3.1 and pCMV /Bsd generate the antibiotic resistance gene cassette by using Polymerase Chain Reaction (PCR) amplification together with 0.2µ1 Phusion DNA polymerase (1 U/50µ1). All PCR products were purified and analyzed with 2.0 % agarose gel electrophoresis method by running alongside with 100bp DNA Ladder Marker and resulting in successful generation of the gene knockout antibiotic resistance gene (GKO-AbR) cassette. It is estimated that these cassettes could be used for further study which are to bind with the CLDN-1 homology arms (left and right), to construct the CLDN-1 knockout plasmids and to be used in gene knockout targeting work for CLDN-1 in HCV permissive cells. 2013 Thesis NonPeerReviewed text en https://ir.uitm.edu.my/id/eprint/109158/1/109158.PDF Generation of gene knockout cassette (GKO-AbR) for claudin-1 gene targeting / Laily Norsafira Sahizan. (2013) Degree thesis, thesis, Universiti Teknologi MARA (Kampus Puncak Alam).
institution Universiti Teknologi Mara
building Tun Abdul Razak Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Mara
content_source UiTM Institutional Repository
url_provider http://ir.uitm.edu.my/
language English
topic Pharmacopoeias
spellingShingle Pharmacopoeias
Sahizan, Laily Norsafira
Generation of gene knockout cassette (GKO-AbR) for claudin-1 gene targeting / Laily Norsafira Sahizan
description Hepatitis C is a type of inflammation of the liver, origin of Hepatitis C Virus (HCV) and it is estimated that more than 170 million people worldwide have been infected with this virus. Claudin-I (CLDN-1) is one of the receptors that are believed to be crucial for HCV entry process, but the details on the mechanisms are still unclear. This research aims to design the primers and amplify the antibiotic resistance gene of interest from the commercially available plasmid and to produce gene knockout cassette of claudin-I by linking the homology arms of exon 1 claudin-I region. PrimerBlast online software was used as the tool to design the target-specific primers ofpcDNA3.l and pCMV/Bsd vector. The gene of interest used from the pcDNA3.1 vector was Neomycin resistance gene while from pCMV/Bsd vector the gene of interest used was Blasticidin resistance gene. The sizes of the primers produced for pcDN A3.1 and pCMV /Bsd generate the antibiotic resistance gene cassette by using Polymerase Chain Reaction (PCR) amplification together with 0.2µ1 Phusion DNA polymerase (1 U/50µ1). All PCR products were purified and analyzed with 2.0 % agarose gel electrophoresis method by running alongside with 100bp DNA Ladder Marker and resulting in successful generation of the gene knockout antibiotic resistance gene (GKO-AbR) cassette. It is estimated that these cassettes could be used for further study which are to bind with the CLDN-1 homology arms (left and right), to construct the CLDN-1 knockout plasmids and to be used in gene knockout targeting work for CLDN-1 in HCV permissive cells.
format Thesis
author Sahizan, Laily Norsafira
author_facet Sahizan, Laily Norsafira
author_sort Sahizan, Laily Norsafira
title Generation of gene knockout cassette (GKO-AbR) for claudin-1 gene targeting / Laily Norsafira Sahizan
title_short Generation of gene knockout cassette (GKO-AbR) for claudin-1 gene targeting / Laily Norsafira Sahizan
title_full Generation of gene knockout cassette (GKO-AbR) for claudin-1 gene targeting / Laily Norsafira Sahizan
title_fullStr Generation of gene knockout cassette (GKO-AbR) for claudin-1 gene targeting / Laily Norsafira Sahizan
title_full_unstemmed Generation of gene knockout cassette (GKO-AbR) for claudin-1 gene targeting / Laily Norsafira Sahizan
title_sort generation of gene knockout cassette (gko-abr) for claudin-1 gene targeting / laily norsafira sahizan
publishDate 2013
url https://ir.uitm.edu.my/id/eprint/109158/1/109158.PDF
https://ir.uitm.edu.my/id/eprint/109158/
_version_ 1825165107555991552
score 13.239859

Similar Items