Evaluation of Antifungal Activity of Defensin (Tfgd2) Using Its Heterologous Expression in E. coli

Defensins are a superfamily of antimicrobial peptides that can inhibit the growth of a broad spectrum of fungi. To evaluate the antifungal activity of Tfgd2 gene of Trigonella foenum graecum, the coding region of this gene in the cDNA form was subcloned in the expression vector pET26b (+) and the co...

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Main Authors: Shiva Shiehbeiki, Mostafa Motallebi, Mohammad Zamani, Esmat Jourabchi
Format: Article
Language:English
Published: University of Mazandaran 2021-02-01
Series:Journal of Genetic Resources
Subjects:
Online Access:http://sc.journals.umz.ac.ir/article_3114_43897496dc534646c7199480c6253f97.pdf
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spelling doaj-06f1ec3d1a274c3f9674fc19679a08ab2021-08-18T09:19:15ZengUniversity of MazandaranJournal of Genetic Resources2423-42572588-25892021-02-017111612410.22080/jgr.2021.20205.12273114Evaluation of Antifungal Activity of Defensin (Tfgd2) Using Its Heterologous Expression in E. coliShiva Shiehbeiki0Mostafa Motallebi1Mohammad Zamani2Esmat Jourabchi3Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.Defensins are a superfamily of antimicrobial peptides that can inhibit the growth of a broad spectrum of fungi. To evaluate the antifungal activity of Tfgd2 gene of Trigonella foenum graecum, the coding region of this gene in the cDNA form was subcloned in the expression vector pET26b (+) and the construct was designated as pETSH1. The obtained construct expressing the recombinant protein with a hexahistidine tag at the C-terminal end transformed into the E. coli BL21 (DE3). Taguchi test applied for optimizing the protein expression and the expressed protein TFGD2 confirmed by SDS-PAGE and western blotting. The recombinant TFGD2 protein was purified using affinity chromatography with a Ni-NTA column. In vitro assay indicated a broad spectrum of antifungal activity of purified expressed recombinant TFGD2 against different fungal phytopathogens, such as Fusarium oxysporum, Rhizoctonia solani, Sclerotinia sclerotiorum, Alternaria solani, and Verticillium dahliae.http://sc.journals.umz.ac.ir/article_3114_43897496dc534646c7199480c6253f97.pdfantifungal activitydefensinsytox greentfgd2pathogenesis-related protein
collection DOAJ
language English
format Article
sources DOAJ
author Shiva Shiehbeiki
Mostafa Motallebi
Mohammad Zamani
Esmat Jourabchi
spellingShingle Shiva Shiehbeiki
Mostafa Motallebi
Mohammad Zamani
Esmat Jourabchi
Evaluation of Antifungal Activity of Defensin (Tfgd2) Using Its Heterologous Expression in E. coli
Journal of Genetic Resources
antifungal activity
defensin
sytox green
tfgd2
pathogenesis-related protein
author_facet Shiva Shiehbeiki
Mostafa Motallebi
Mohammad Zamani
Esmat Jourabchi
author_sort Shiva Shiehbeiki
title Evaluation of Antifungal Activity of Defensin (Tfgd2) Using Its Heterologous Expression in E. coli
title_short Evaluation of Antifungal Activity of Defensin (Tfgd2) Using Its Heterologous Expression in E. coli
title_full Evaluation of Antifungal Activity of Defensin (Tfgd2) Using Its Heterologous Expression in E. coli
title_fullStr Evaluation of Antifungal Activity of Defensin (Tfgd2) Using Its Heterologous Expression in E. coli
title_full_unstemmed Evaluation of Antifungal Activity of Defensin (Tfgd2) Using Its Heterologous Expression in E. coli
title_sort evaluation of antifungal activity of defensin (tfgd2) using its heterologous expression in e. coli
publisher University of Mazandaran
series Journal of Genetic Resources
issn 2423-4257
2588-2589
publishDate 2021-02-01
description Defensins are a superfamily of antimicrobial peptides that can inhibit the growth of a broad spectrum of fungi. To evaluate the antifungal activity of Tfgd2 gene of Trigonella foenum graecum, the coding region of this gene in the cDNA form was subcloned in the expression vector pET26b (+) and the construct was designated as pETSH1. The obtained construct expressing the recombinant protein with a hexahistidine tag at the C-terminal end transformed into the E. coli BL21 (DE3). Taguchi test applied for optimizing the protein expression and the expressed protein TFGD2 confirmed by SDS-PAGE and western blotting. The recombinant TFGD2 protein was purified using affinity chromatography with a Ni-NTA column. In vitro assay indicated a broad spectrum of antifungal activity of purified expressed recombinant TFGD2 against different fungal phytopathogens, such as Fusarium oxysporum, Rhizoctonia solani, Sclerotinia sclerotiorum, Alternaria solani, and Verticillium dahliae.
topic antifungal activity
defensin
sytox green
tfgd2
pathogenesis-related protein
url http://sc.journals.umz.ac.ir/article_3114_43897496dc534646c7199480c6253f97.pdf
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