Molecular cloning and production of recombinant phytase from Bacillus Subtilis ASUIA243 in Pichia Pastoris

Molecular cloning and production of recombinant phytase from Bacillus Subtilis ASUIA243 in Pichia Pastoris

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 linea...

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Main Authors: Mohd Dali, Nor Soleha, Nuge, Tamrin, Mahamad Maifiah, Mohd Hafidz, Yusof, Faridah, Meor Hussin, Anis Shobirin, Farouk, Abd Alaziem, Mohd. Salleh, Hamzah
Format: Article
Language:English
Published: IIUM Press 2011
Subjects:
QH426 Genetics
Online Access:http://irep.iium.edu.my/17110/
http://irep.iium.edu.my/17110/
http://irep.iium.edu.my/17110/1/molecular_cloning.pdf
id iium-17110
recordtype eprints
spelling iium-171102012-07-06T02:26:12Z http://irep.iium.edu.my/17110/ Molecular cloning and production of recombinant phytase from Bacillus Subtilis ASUIA243 in Pichia Pastoris Mohd Dali, Nor Soleha Nuge, Tamrin Mahamad Maifiah, Mohd Hafidz Yusof, Faridah Meor Hussin, Anis Shobirin 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/17110/1/molecular_cloning.pdf Mohd Dali, Nor Soleha and Nuge, Tamrin and Mahamad Maifiah, Mohd Hafidz and Yusof, Faridah and Meor Hussin, Anis Shobirin and Farouk, Abd Alaziem and Mohd. Salleh, Hamzah (2011) Molecular cloning and production of recombinant phytase from Bacillus Subtilis ASUIA243 in Pichia Pastoris. IIUM Engineering Journal, 12 (4). p. 1. ISSN 1511-788X http://www.iium.edu.my/ejournal/index.php/iiumej/article/view/211/259
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
Mohd Dali, Nor Soleha
Nuge, Tamrin
Mahamad Maifiah, Mohd Hafidz
Yusof, Faridah
Meor Hussin, Anis Shobirin
Farouk, Abd Alaziem
Mohd. Salleh, Hamzah
Molecular cloning and production of recombinant phytase from Bacillus Subtilis ASUIA243 in Pichia Pastoris
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 Mohd Dali, Nor Soleha
Nuge, Tamrin
Mahamad Maifiah, Mohd Hafidz
Yusof, Faridah
Meor Hussin, Anis Shobirin
Farouk, Abd Alaziem
Mohd. Salleh, Hamzah
author_facet Mohd Dali, Nor Soleha
Nuge, Tamrin
Mahamad Maifiah, Mohd Hafidz
Yusof, Faridah
Meor Hussin, Anis Shobirin
Farouk, Abd Alaziem
Mohd. Salleh, Hamzah
author_sort Mohd Dali, Nor Soleha
title Molecular cloning and production of recombinant phytase from Bacillus Subtilis ASUIA243 in Pichia Pastoris
title_short Molecular cloning and production of recombinant phytase from Bacillus Subtilis ASUIA243 in Pichia Pastoris
title_full Molecular cloning and production of recombinant phytase from Bacillus Subtilis ASUIA243 in Pichia Pastoris
title_fullStr Molecular cloning and production of recombinant phytase from Bacillus Subtilis ASUIA243 in Pichia Pastoris
title_full_unstemmed Molecular cloning and production of recombinant phytase from Bacillus Subtilis ASUIA243 in Pichia Pastoris
title_sort molecular cloning and production of recombinant phytase from bacillus subtilis asuia243 in pichia pastoris
publisher IIUM Press
publishDate 2011
url http://irep.iium.edu.my/17110/
http://irep.iium.edu.my/17110/
http://irep.iium.edu.my/17110/1/molecular_cloning.pdf
first_indexed 2023-09-18T20:25:55Z
last_indexed 2023-09-18T20:25:55Z
_version_ 1777408415105024000

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