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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| Format: | Article |
| Language: | English |
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IIUM Press
2011
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| Online Access: | http://irep.iium.edu.my/17046/ http://irep.iium.edu.my/17046/4/219-1318-1-PB.pdf |
| _version_ | 1848777987671982080 |
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| author | Maifiah, M.H. Mahamad Jamal, Parveen Nuge, Tamrin Mohd Dali, Nor Soleha Meor Hussin, Anis Shobirin Farouk, Abd Alaziem Mohd. Salleh, Hamzah |
| author_facet | Maifiah, M.H. Mahamad Jamal, Parveen Nuge, Tamrin Mohd Dali, Nor Soleha Meor Hussin, Anis Shobirin Farouk, Abd Alaziem Mohd. Salleh, Hamzah |
| author_sort | Maifiah, M.H. Mahamad |
| building | IIUM Repository |
| collection | Online Access |
| 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. |
| first_indexed | 2025-11-14T14:54:43Z |
| format | Article |
| id | iium-17046 |
| institution | International Islamic University Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T14:54:43Z |
| publishDate | 2011 |
| publisher | IIUM Press |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | iium-170462021-07-06T02:28:39Z 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, Nor Soleha 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/17046/4/219-1318-1-PB.pdf Maifiah, M.H. Mahamad and Jamal, Parveen and Nuge, Tamrin and Mohd Dali, Nor Soleha and Meor Hussin, Anis Shobirin 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 |
| spellingShingle | QH426 Genetics Maifiah, M.H. Mahamad Jamal, Parveen Nuge, Tamrin Mohd Dali, Nor Soleha Meor Hussin, Anis Shobirin Farouk, Abd Alaziem Mohd. Salleh, Hamzah Molecular cloning of phytase gene from ASUIA279 and its expression in Pichia Pastoris system |
| title | 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_short | 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 |
| topic | QH426 Genetics |
| 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 |