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: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English English |
| Published: |
2011
|
| Online Access: | http://psasir.upm.edu.my/id/eprint/24206/1/Molecular%20cloning%20and%20production%20of%20recombinant%20phytase%20from%20bacillus%20subtilis%20ASUIA243%20in%20Pichia%20Pastoris.pdf |
| Summary: | 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. |
|---|