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

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

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Abstract

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.

Item Type: Article (Journal)
Additional Information: 1813/17046
Uncontrolled Keywords: phytase; Bacillus subtilis; Pichia pastoris; gene cloning
Subjects: Q Science > QH Natural history > QH426 Genetics
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button): Kulliyyah of Engineering > Department of Biotechnology Engineering
Depositing User: Mrs Maznah Zakaria
Date Deposited: 26 Jan 2012 09:06
Last Modified: 06 Mar 2012 17:22
URI: http://irep.iium.edu.my/id/eprint/17046

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