Open AccessCloning of Bacillus subtilis phytase gene construct in Escherichia coli
Author(s) -
Mahdiyar Iravani Saadi,
Abbas Doosti,
Heeva Jalali,
Ehsan Nabi Abdolyousefi,
Mansooreh Hooshiyar,
Reza Tabrizi,
Esmat Noshadi
Publication year - 2021
Publication title -
iranian journal of microbiology.
Language(s) - English
Resource type - Journals
eISSN - 2008-4447
pISSN - 2008-3289
DOI - 10.18502/ijm.v13i5.7433
Subject(s) - phytase , bacillus subtilis , escherichia coli , biology , phytic acid , microbiology and biotechnology , gene , cloning (programming) , molecular cloning , biochemistry , gene expression , bacteria , enzyme , genetics , computer science , programming language
Background and Objectives: Phytase has a hydrolysis function of phytic acid, which yields inorganic phosphate. Bacillus species can produce thermostable alkaline phytase. The aim of this study was to isolate and clone a Phytase gene (Phy) from Bacillus subtilis in Escherichia coli.
Materials and Methods: In this study, the extracellular PhyC gene was isolated from Bacillus subtilis Phytase C. After purification of the bands, DNA fragment of Phy gene was cloned by T/A cloning technique, and the clone was transformed into Escherichia coli. Afterward, the pGEM-Phy was transferred into E. coli Top-10 strain and the recombinants were plated on LB agar containing 100 µg/ml ampicillin. The colonization of 1171 bp of gene Phytase C was confirmed by PCR. The presence of gene-targeting in vector was confirmed with enzymatic digestion by XhoI and XbaI restriction enzymes.
Results: The Phytase gene was successfully cloned in E. coli. The result of cloning of 1171 bp Phytase gene was confirmed by PCR assay.
Conclusion: Our impression of this article is that several methods, such as using along with microbial, plant phytase reproduction, or low-phytic acid corn may be the better way from a single phytase.