Cloning of Bacillus subtilis phytase gene construct in Escherichia coli
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 Met...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Tehran University of Medical Sciences
2021-10-01
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| Series: | Iranian Journal of Microbiology |
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| Online Access: | https://ijm.tums.ac.ir/index.php/ijm/article/view/3283 |
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| author | Mahdiyar Iravani Saadi Abbas Doosti Heeva Jalali Ehsan Nabi Abdolyousefi Mansooreh Hooshiyar Reza Tabrizi Esmat Noshadi |
| author_facet | Mahdiyar Iravani Saadi Abbas Doosti Heeva Jalali Ehsan Nabi Abdolyousefi Mansooreh Hooshiyar Reza Tabrizi Esmat Noshadi |
| author_sort | Mahdiyar Iravani Saadi |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-14T02:57:01Z |
| format | Article |
| id | doaj.art-98de0de4d1a240d7b053ad9536fdb8b5 |
| institution | Directory Open Access Journal |
| issn | 2008-3289 2008-4447 |
| language | English |
| last_indexed | 2024-12-14T02:57:01Z |
| publishDate | 2021-10-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Microbiology |
| spelling | doaj.art-98de0de4d1a240d7b053ad9536fdb8b52022-12-21T23:19:35ZengTehran University of Medical SciencesIranian Journal of Microbiology2008-32892008-44472021-10-0113510.18502/ijm.v13i5.7433Cloning of Bacillus subtilis phytase gene construct in Escherichia coliMahdiyar Iravani Saadi0Abbas Doosti1Heeva Jalali2Ehsan Nabi Abdolyousefi3Mansooreh Hooshiyar4Reza Tabrizi5Esmat Noshadi6Hematology Research Center, Shiraz University of Medical Sciences, Shiraz, IranBiotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, IranDepartment of Animal Science, Faculty of Agriculture, University of Kurdistan, Sanandaj, IranHematology Research Center, Shiraz University of Medical Sciences, Shiraz, IranDepartment of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, IranNon-Communicable Diseases Research Center, Fasa University of Medical Sciences, Fasa, IranHematology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran; Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, IranBackground 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.https://ijm.tums.ac.ir/index.php/ijm/article/view/3283Bacillus subtilis;Cloning;Escherichia coli;Phytase;Probiotics |
| spellingShingle | Mahdiyar Iravani Saadi Abbas Doosti Heeva Jalali Ehsan Nabi Abdolyousefi Mansooreh Hooshiyar Reza Tabrizi Esmat Noshadi Cloning of Bacillus subtilis phytase gene construct in Escherichia coli Iranian Journal of Microbiology Bacillus subtilis; Cloning; Escherichia coli; Phytase; Probiotics |
| title | Cloning of Bacillus subtilis phytase gene construct in Escherichia coli |
| title_full | Cloning of Bacillus subtilis phytase gene construct in Escherichia coli |
| title_fullStr | Cloning of Bacillus subtilis phytase gene construct in Escherichia coli |
| title_full_unstemmed | Cloning of Bacillus subtilis phytase gene construct in Escherichia coli |
| title_short | Cloning of Bacillus subtilis phytase gene construct in Escherichia coli |
| title_sort | cloning of bacillus subtilis phytase gene construct in escherichia coli |
| topic | Bacillus subtilis; Cloning; Escherichia coli; Phytase; Probiotics |
| url | https://ijm.tums.ac.ir/index.php/ijm/article/view/3283 |
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