Author/Authors :
DALI, NOR SOLEHA MOHD International Islamic University Malaysia - Bioprocess and Molecular Engineering Research Unit,Department of Biotechnology Engineering, Malaysia , NUGE, TAMRIN International Islamic University Malaysia - Bioprocess and Molecular Engineering Research Unit,Department of Biotechnology Engineering, Malaysia , MAIFIAH, MOHD. HAFIDZ MAHAMAD International Islamic University Malaysia - Bioprocess and Molecular Engineering Research Unit,Department of Biotechnology Engineering, Malaysia , YUSOF, FARIDAH International Islamic University Malaysia - Bioprocess and Molecular Engineering Research Unit,Department of Biotechnology Engineering, Malaysia , HUSSIN, ANIS SHOBIRIN MEOR Universiti Putra Malaysia - Faculty of Science and Food Technology - Department of Food Technology, Malaysia , FAROUK, ABD-ELAZIEM Taif University - Faculty of Science - Department of Biotechnology, Saudi Arabia , SALLEH, HAMZAH MOHD. International Islamic University Malaysia - Bioprocess and Molecular Engineering Research Unit,Department of Biotechnology Engineering, Malaysia
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.
Keywords :
phytase , Bacillus subtilis , Pichia pastoris , gene cloning
