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

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

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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: 4734/17110
Uncontrolled Keywords: phytase; Bacillus subtilis; Pichia pastoris; gene cloning
Subjects: Q Science > QH Natural history > QH426 Genetics
Kulliyyahs/Centres/Divisions/Institutes: Kulliyyah of Engineering > Department of Biotechnology Engineering
Depositing User: DR FARIDAH YUSOF
Date Deposited: 26 Jan 2012 14:54
Last Modified: 06 Jul 2012 10:26
URI: http://irep.iium.edu.my/id/eprint/17110

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