Cloning of human gamma glutamyl transferase (GGT) gene / Izzati Mohd Farok

The y-glutamyl transferase (GGT) plays an important role in the process of catalyzing the degradation of glutathione. GGT is widely used as a marker in preneoplastic lesions in the liver during chemical carcinogenesis. In this study, primers were designed without the restriction sites to investigate...

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Main Author:	Mohd Farok, Izzati
Format:	Thesis
Language:	English
Published:	2008
Subjects:	DNA Deoxyribonucleic acids Cloning Genomics
Online Access:	https://ir.uitm.edu.my/id/eprint/101083/1/101083.PDF
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id	my-uitm-ir.101083
record_format	uketd_dc
spelling	my-uitm-ir.1010832024-08-28T18:54:15Z Cloning of human gamma glutamyl transferase (GGT) gene / Izzati Mohd Farok 2008 Mohd Farok, Izzati DNA. Deoxyribonucleic acids Cloning Genomics The y-glutamyl transferase (GGT) plays an important role in the process of catalyzing the degradation of glutathione. GGT is widely used as a marker in preneoplastic lesions in the liver during chemical carcinogenesis. In this study, primers were designed without the restriction sites to investigate the success of cloning. The aim of this study was to clone GGT gene in plasmid by means of TA cloning. Methods used in this study were purification of human liver cDNA, PCR for gene amplification, TOPO TA cloning, transformation into E.coli and DNA sequencing. PCR was conducted under different annealing temperatures using Takara PCR Thermal Cycler Dice™ Gradient model to determine the optimal annealing temperature. TOPO TA cloning process with high cloning efficiency was completed in 5 minutes. Transformation was done using the One Shot® Chemical Competent E.coli. DNA sequencing was also performed using the plasmid samples from transformation process to validate the insert. Plasmid extraction yielded 168 ng/µl of liver cDNA. The PCR based method obtained an optimal annealing temperature for GGT amplification, which was at 47°C. Transformation was a success by using E.coli as a host where a 10 ml of LB broth was used and incubated at 3 7°C for 16 hours. Cloning of the GGT gene is important as it opens ways for further studies to look into its association with cancer and help in development of medication that prevents the progression of tumors. There is no specific treatment for patients with GGT deficiency and thus, researchers may develop protein-based drugs for them. GGT cloning can help in the establishment of gene collection in DNA libraries. 2008 Thesis https://ir.uitm.edu.my/id/eprint/101083/ https://ir.uitm.edu.my/id/eprint/101083/1/101083.PDF text en public degree Universiti Teknologi MARA (Kampus Puncak Alam) Faculty of Pharmacy Kek, Teh Lay
institution	Universiti Teknologi MARA
collection	UiTM Institutional Repository
language	English
advisor	Kek, Teh Lay
topic	DNA Deoxyribonucleic acids Cloning Genomics
spellingShingle	DNA Deoxyribonucleic acids Cloning Genomics Mohd Farok, Izzati Cloning of human gamma glutamyl transferase (GGT) gene / Izzati Mohd Farok
description	The y-glutamyl transferase (GGT) plays an important role in the process of catalyzing the degradation of glutathione. GGT is widely used as a marker in preneoplastic lesions in the liver during chemical carcinogenesis. In this study, primers were designed without the restriction sites to investigate the success of cloning. The aim of this study was to clone GGT gene in plasmid by means of TA cloning. Methods used in this study were purification of human liver cDNA, PCR for gene amplification, TOPO TA cloning, transformation into E.coli and DNA sequencing. PCR was conducted under different annealing temperatures using Takara PCR Thermal Cycler Dice™ Gradient model to determine the optimal annealing temperature. TOPO TA cloning process with high cloning efficiency was completed in 5 minutes. Transformation was done using the One Shot® Chemical Competent E.coli. DNA sequencing was also performed using the plasmid samples from transformation process to validate the insert. Plasmid extraction yielded 168 ng/µl of liver cDNA. The PCR based method obtained an optimal annealing temperature for GGT amplification, which was at 47°C. Transformation was a success by using E.coli as a host where a 10 ml of LB broth was used and incubated at 3 7°C for 16 hours. Cloning of the GGT gene is important as it opens ways for further studies to look into its association with cancer and help in development of medication that prevents the progression of tumors. There is no specific treatment for patients with GGT deficiency and thus, researchers may develop protein-based drugs for them. GGT cloning can help in the establishment of gene collection in DNA libraries.
format	Thesis
qualification_level	Bachelor degree
author	Mohd Farok, Izzati
author_facet	Mohd Farok, Izzati
author_sort	Mohd Farok, Izzati
title	Cloning of human gamma glutamyl transferase (GGT) gene / Izzati Mohd Farok
title_short	Cloning of human gamma glutamyl transferase (GGT) gene / Izzati Mohd Farok
title_full	Cloning of human gamma glutamyl transferase (GGT) gene / Izzati Mohd Farok
title_fullStr	Cloning of human gamma glutamyl transferase (GGT) gene / Izzati Mohd Farok
title_full_unstemmed	Cloning of human gamma glutamyl transferase (GGT) gene / Izzati Mohd Farok
title_sort	cloning of human gamma glutamyl transferase (ggt) gene / izzati mohd farok
granting_institution	Universiti Teknologi MARA (Kampus Puncak Alam)
granting_department	Faculty of Pharmacy
publishDate	2008
url	https://ir.uitm.edu.my/id/eprint/101083/1/101083.PDF
_version_	1811769127728054272

Cloning of human gamma glutamyl transferase (GGT) gene / Izzati Mohd Farok

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