Molecular cloning of phytase gene from ASUIA279 and its expression in Pichia Pastoris system

Phytase gene obtained from Bacillus subtilis ASUIA243 was cloned into a medium vector and transformed into E. coli. Restriction enzyme digestion was conducted to get blunt-ended phytase gene and ligated into the Pichia expression vector, pPICZαA. The recombinant vector, pPICZαA-243HPp was then li...

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Main Authors:	Maifiah, M.H. Mahamad, Jamal, Parveen, Nuge, Tamrin, Mohd Dali, N.S., Meor Hussin, A.S., Farouk, Abd Alaziem, Mohd. Salleh, Hamzah
Format:	Article
Language:	English
Published:	IIUM Press 2011
Subjects:	QH426 Genetics
Online Access:	http://irep.iium.edu.my/17046/ http://irep.iium.edu.my/17046/ http://irep.iium.edu.my/17046/4/219-1318-1-PB.pdf

id	iium-17046
recordtype	eprints
spelling	iium-170462012-03-06T09:22:01Z http://irep.iium.edu.my/17046/ Molecular cloning of phytase gene from ASUIA279 and its expression in Pichia Pastoris system Maifiah, M.H. Mahamad Jamal, Parveen Nuge, Tamrin Mohd Dali, N.S. Meor Hussin, A.S. Farouk, Abd Alaziem Mohd. Salleh, Hamzah QH426 Genetics Phytase gene obtained from Bacillus subtilis ASUIA243 was cloned into a medium vector and transformed into E. coli. Restriction enzyme digestion was conducted to get blunt-ended phytase gene and ligated into the Pichia expression vector, pPICZαA. The recombinant vector, pPICZαA-243HPp was then linearized with PmeI and transformed into P. pastoris strain X33. Screening for multi copy gene number of transformants was done by re-plating the selected colonies on increasing concentration of zeocin. One positive clone, X243HPp#2 was then grown in BMGY media as the starting culture, followed by induction in BMMY media for protein expression study. The supernatant was then analysed by SDS-PAGE and Western blot method to check the protein expression. IIUM Press 2011 Article PeerReviewed application/pdf en http://irep.iium.edu.my/17046/4/219-1318-1-PB.pdf Maifiah, M.H. Mahamad and Jamal, Parveen and Nuge, Tamrin and Mohd Dali, N.S. and Meor Hussin, A.S. and Farouk, Abd Alaziem and Mohd. Salleh, Hamzah (2011) Molecular cloning of phytase gene from ASUIA279 and its expression in Pichia Pastoris system. IIUM Engineering Journal, 12 (4). pp. 1-8. ISSN 1511-788X http://www.iium.edu.my/ejournal/index.php/iiumej/article/view/211
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	QH426 Genetics
spellingShingle	QH426 Genetics Maifiah, M.H. Mahamad Jamal, Parveen Nuge, Tamrin Mohd Dali, N.S. Meor Hussin, A.S. Farouk, Abd Alaziem Mohd. Salleh, Hamzah Molecular cloning of phytase gene from ASUIA279 and its expression in Pichia Pastoris system
description	Phytase gene obtained from Bacillus subtilis ASUIA243 was cloned into a medium vector and transformed into E. coli. Restriction enzyme digestion was conducted to get blunt-ended phytase gene and ligated into the Pichia expression vector, pPICZαA. The recombinant vector, pPICZαA-243HPp was then linearized with PmeI and transformed into P. pastoris strain X33. Screening for multi copy gene number of transformants was done by re-plating the selected colonies on increasing concentration of zeocin. One positive clone, X243HPp#2 was then grown in BMGY media as the starting culture, followed by induction in BMMY media for protein expression study. The supernatant was then analysed by SDS-PAGE and Western blot method to check the protein expression.
format	Article
author	Maifiah, M.H. Mahamad Jamal, Parveen Nuge, Tamrin Mohd Dali, N.S. Meor Hussin, A.S. Farouk, Abd Alaziem Mohd. Salleh, Hamzah
author_facet	Maifiah, M.H. Mahamad Jamal, Parveen Nuge, Tamrin Mohd Dali, N.S. Meor Hussin, A.S. Farouk, Abd Alaziem Mohd. Salleh, Hamzah
author_sort	Maifiah, M.H. Mahamad
title	Molecular cloning of phytase gene from ASUIA279 and its expression in Pichia Pastoris system
title_short	Molecular cloning of phytase gene from ASUIA279 and its expression in Pichia Pastoris system
title_full	Molecular cloning of phytase gene from ASUIA279 and its expression in Pichia Pastoris system
title_fullStr	Molecular cloning of phytase gene from ASUIA279 and its expression in Pichia Pastoris system
title_full_unstemmed	Molecular cloning of phytase gene from ASUIA279 and its expression in Pichia Pastoris system
title_sort	molecular cloning of phytase gene from asuia279 and its expression in pichia pastoris system
publisher	IIUM Press
publishDate	2011
url	http://irep.iium.edu.my/17046/ http://irep.iium.edu.my/17046/ http://irep.iium.edu.my/17046/4/219-1318-1-PB.pdf
first_indexed	2023-09-18T20:25:51Z
last_indexed	2023-09-18T20:25:51Z
_version_	1777408411628994560

Molecular cloning of phytase gene from ASUIA279 and its expression in Pichia Pastoris system

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