DESIGN, SYNTHESIS, CLONING, EXPRESSION AND EVALUATION OF THE ANTIBACTERIAL EFFECT OF WHEAT DEFENSIN (TRITICUM AESTIVUM) IN E.COLI HOST

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MORTAZAVI S.M.; SAFFAR B.; MIRAKHORLI N.; MOBINI DEHKORDI M.

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Journal: Journal of Molecular and Cellular Researches Year:2016 | Volume:29 | Issue:1 Page(s): 114-125

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Title

DESIGN, SYNTHESIS, CLONING, EXPRESSION AND EVALUATION OF THE ANTIBACTERIAL EFFECT OF WHEAT DEFENSIN (TRITICUM AESTIVUM) IN E.COLI HOST

Author(s)

MORTAZAVI S.M. | SAFFAR B. | MIRAKHORLI N. | MOBINI DEHKORDI M. | Issue Writer Certificate

Keywords

ANTIMICROBIAL PEPTIDESQ1

WHEAT DEFENSIN PROTEINQ1

CLONING AND EXPRESSIONQ1

Abstract

Plant defensins are a family of cationic ANTIMICROBIAL PEPTIDES, small and cysteine rich that have a broad range of antibacterial effects. The aim of this study was designed, synthesis, cloning, expression of wheat defensin (Triticum aestivum) gene and evaluation of the antibacterial effect of recombinant protein. The defensin gene was fused to sumo gene for more expression and solubility and then was optimized by some software and was synthesized and cloned in pGH vector. Then sumo_defensin sequence was sub cloned into pET-32a (+) vector and transformed into bacterial cells (E.coli origami). Gene expression was induced by 1mM IPTG in 30oC and proteins were analyzed by SDS-PAGE and Western blotting. The fusion protein with a molecular weight of approximately 38 kDa was analyzed on SDS-PAGE gel and confirmed by western blotting. After digestion of protein by Sumo protease, the antibacterial effect of the recombinant wheat defensin was evaluated on the plant pathogenic Gram-negative bacteria Xanthomonase axonopodis, Pseudomonas syringae and some human pathogenic Gram-negative and Gram-positive bacteria such as E.coli and Klebsiella pneumoniae , Staphylococcus aureus and Bacillus cereus. In the end, recombinant wheat defensin had an antibacterial effect against Xanthomonase axonopodis, Staphylococcus aureus and Bacillus cereus. Based on the antimicrobial characteristics of wheat defensin, this study can be effective to introduce a new antimicrobial agent.

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APA: Copy

MORTAZAVI, S.M., SAFFAR, B., MIRAKHORLI, N., & MOBINI DEHKORDI, M.. (2016). DESIGN, SYNTHESIS, CLONING, EXPRESSION AND EVALUATION OF THE ANTIBACTERIAL EFFECT OF WHEAT DEFENSIN (TRITICUM AESTIVUM) IN E.COLI HOST. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), 29(1 ), 114-125. SID. https://sid.ir/paper/248435/en

Vancouver: Copy

MORTAZAVI S.M., SAFFAR B., MIRAKHORLI N., MOBINI DEHKORDI M.. DESIGN, SYNTHESIS, CLONING, EXPRESSION AND EVALUATION OF THE ANTIBACTERIAL EFFECT OF WHEAT DEFENSIN (TRITICUM AESTIVUM) IN E.COLI HOST. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY)[Internet]. 2016;29(1 ):114-125. Available from: https://sid.ir/paper/248435/en

IEEE: Copy

S.M. MORTAZAVI, B. SAFFAR, N. MIRAKHORLI, and M. MOBINI DEHKORDI, “DESIGN, SYNTHESIS, CLONING, EXPRESSION AND EVALUATION OF THE ANTIBACTERIAL EFFECT OF WHEAT DEFENSIN (TRITICUM AESTIVUM) IN E.COLI HOST,” JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), vol. 29, no. 1 , pp. 114–125, 2016, [Online]. Available: https://sid.ir/paper/248435/en

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