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...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
IIUM Press
2011
|
| Subjects: | |
| Online Access: | http://irep.iium.edu.my/17110/ http://irep.iium.edu.my/17110/1/molecular_cloning.pdf |
| _version_ | 1848777994380771328 |
|---|---|
| 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 |
| 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 | 2025-11-14T14:54:49Z |
| format | Article |
| id | iium-17110 |
| institution | International Islamic University Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T14:54:49Z |
| publishDate | 2011 |
| publisher | IIUM Press |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | iium-171102021-07-06T02:30:40Z 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 |
| 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 |
| title | 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_short | 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 |
| topic | QH426 Genetics |
| url | http://irep.iium.edu.my/17110/ http://irep.iium.edu.my/17110/ http://irep.iium.edu.my/17110/1/molecular_cloning.pdf |