MOLECULAR CLONING AND PRODUCTION OF RECOMBINANT PHYTASE FROM BACILLUS SUBTILIS ASUIA243 IN PICHIA PASTORIS

被引：0

作者：

Dali, Nor Soleha Mohd ^{[1
]}

Nuge, Tamrin ^{[1
]}

Maifiah, Mohd. Hafidz Mahamad ^{[1
]}

Yusof, Faridah ^{[1
]}

Hussin, Anis Shobirin Meor ^{[2
]}

Farouk, Abd-Elaziem ^{[3
]}

Salleh, Hamzah Mohd. ^{[1
]}

机构：

[1] Int Islamic Univ Malaysia, Dept Biotechnol Engn, Bioproc & Mol Engn Res Unit, Kuala Lumpur 53100, Malaysia

[2] Univ Putra Malaysia, Fac Sci & Food Technol, Dept Food Technol, Serdang 43400, Selangor, Malaysia

[3] Taif Univ, Fac Sci, Dept Biotechnol, At Taif 21974, Al Hawiayah, Saudi Arabia

来源：

IIUM ENGINEERING JOURNAL | 2011年 / 12卷 / 04期

关键词：

phytase; Bacillus subtilis; Pichia pastoris; gene cloning;

D O I：

暂无

中图分类号：

T [工业技术];

学科分类号：

08 ;

摘要：

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 alpha A. The recombinant vector, pPICZ alpha 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.

引用

页码：99 / 108

页数：10

共 33 条

[1] SUMO fusion technology for difficult-to-express proteins [J].