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, pPICZ�A. The recombinant vector, pPICZ�A-243HPp was then linea...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English English |
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2011
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| Online Access: | http://psasir.upm.edu.my/id/eprint/24206/ http://psasir.upm.edu.my/id/eprint/24206/1/Molecular%20cloning%20and%20production%20of%20recombinant%20phytase%20from%20bacillus%20subtilis%20ASUIA243%20in%20Pichia%20Pastoris.pdf |
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| author | Mohd Dali , Nor Soleha Nuge , Tamrin Mahamad Maifiah, Mohd. Hafidz Yusof, Faridah Meor Hussin, Anis Shobirin Abd. El Aziem, Farouk Mohd. Salleh, Hamzah |
| author_facet | Mohd Dali , Nor Soleha Nuge , Tamrin Mahamad Maifiah, Mohd. Hafidz Yusof, Faridah Meor Hussin, Anis Shobirin Abd. El Aziem, Farouk Mohd. Salleh, Hamzah |
| author_sort | Mohd Dali , Nor Soleha |
| building | UPM Institutional 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, pPICZ�A. The recombinant vector, pPICZ�A-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 | 2025-11-15T08:39:24Z |
| format | Article |
| id | upm-24206 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T08:39:24Z |
| publishDate | 2011 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-242062015-10-20T00:35:14Z http://psasir.upm.edu.my/id/eprint/24206/ 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 Abd. El Aziem, Farouk Mohd. Salleh, Hamzah 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, pPICZ�A. The recombinant vector, pPICZ�A-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. 2011 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/24206/1/Molecular%20cloning%20and%20production%20of%20recombinant%20phytase%20from%20bacillus%20subtilis%20ASUIA243%20in%20Pichia%20Pastoris.pdf Mohd Dali , Nor Soleha and Nuge , Tamrin and Mahamad Maifiah, Mohd. Hafidz and Yusof, Faridah and Meor Hussin, Anis Shobirin and Abd. El Aziem, Farouk and Mohd. Salleh, Hamzah (2011) Molecular cloning and production of recombinant phytase from bacillus subtilis ASUIA243 in Pichia Pastoris. IIUM Engineering Journal, 12 (4). pp. 99-108. ISSN 1511-788X English |
| spellingShingle | Mohd Dali , Nor Soleha Nuge , Tamrin Mahamad Maifiah, Mohd. Hafidz Yusof, Faridah Meor Hussin, Anis Shobirin Abd. El Aziem, Farouk Mohd. Salleh, Hamzah Molecular cloning and production of recombinant phytase from bacillus subtilis ASUIA243 in Pichia Pastoris. |
| title | Molecular cloning and production of recombinant phytase from bacillus subtilis ASUIA243 in Pichia Pastoris.
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| title_full | Molecular cloning and production of recombinant phytase from bacillus subtilis ASUIA243 in Pichia Pastoris.
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| 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.
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| title_short | Molecular cloning and production of recombinant phytase from bacillus subtilis ASUIA243 in Pichia Pastoris.
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| title_sort | molecular cloning and production of recombinant phytase from bacillus subtilis asuia243 in pichia pastoris. |
| url | http://psasir.upm.edu.my/id/eprint/24206/ http://psasir.upm.edu.my/id/eprint/24206/1/Molecular%20cloning%20and%20production%20of%20recombinant%20phytase%20from%20bacillus%20subtilis%20ASUIA243%20in%20Pichia%20Pastoris.pdf |