Mohamed, S., El-Mahdy, M., Mabrouk, A., Salem, R., Shehata, M., El-Kholy, I., Abouseadaa, H. (2023). Molecular Characterization, Heterologous Expression and Antimicrobial Activity of Phaseolus vulgaris L. Defensin Peptide (Pv-Def) against various Human MDR Pathogens. Egyptian Journal of Botany, 63(3), 841-849. doi: 10.21608/ejbo.2023.199400.2277
Shereen S. Mohamed; Mohamed M. El-Mahdy; Alshaimaa M. Mabrouk; Reda Salem; Maher M. Shehata; Iman M.A. El-Kholy; Heba H. Abouseadaa. "Molecular Characterization, Heterologous Expression and Antimicrobial Activity of Phaseolus vulgaris L. Defensin Peptide (Pv-Def) against various Human MDR Pathogens". Egyptian Journal of Botany, 63, 3, 2023, 841-849. doi: 10.21608/ejbo.2023.199400.2277
Mohamed, S., El-Mahdy, M., Mabrouk, A., Salem, R., Shehata, M., El-Kholy, I., Abouseadaa, H. (2023). 'Molecular Characterization, Heterologous Expression and Antimicrobial Activity of Phaseolus vulgaris L. Defensin Peptide (Pv-Def) against various Human MDR Pathogens', Egyptian Journal of Botany, 63(3), pp. 841-849. doi: 10.21608/ejbo.2023.199400.2277
Mohamed, S., El-Mahdy, M., Mabrouk, A., Salem, R., Shehata, M., El-Kholy, I., Abouseadaa, H. Molecular Characterization, Heterologous Expression and Antimicrobial Activity of Phaseolus vulgaris L. Defensin Peptide (Pv-Def) against various Human MDR Pathogens. Egyptian Journal of Botany, 2023; 63(3): 841-849. doi: 10.21608/ejbo.2023.199400.2277
Molecular Characterization, Heterologous Expression and Antimicrobial Activity of Phaseolus vulgaris L. Defensin Peptide (Pv-Def) against various Human MDR Pathogens
1Ain Shams Specialized Hospital, Ain Shams University, Cairo, Egypt
2Department of Bioinformatics, Agricultural Genetic Engineering Research Institute, Agricultural Research Center, Giza, Egypt
3Department of Botany, Faculty of Science, Ain Shams University, Cairo, Egypt
Abstract
The elevating worldwide threatening antibiotic resistance, without effective antimicrobials for the prevention and treatment of infections, has put human health at very high risk. Plant defensins are cysteine -rich small cationic peptides, whose tertiary structure is greatly stable due to the presence of disulfide bonds. They exhibit significant antimicrobial activities. The defensin cDNA was successfully obtained from the total RNA extracted from Phaseolusvulgaris L. (Cv. Paulista) young green leaves, the CDS was efficiently cloned into pJET1.2/blunt cloning vector, and then subcloned into the prokaryotic expression vector pGEX -4 T -1. To facilitate defensin peptide purification, the sequence fragment was fused in -frame with a GST tag. The pGEX -4 T -1 containing recombinant defensin (pGEX -4 T - 1 -Pv -Def) was transformed into the E. coli strain BL21. Glutathione affinity chromatography method of purification yielded 3.7 mg/L of recombinant peptide. Western blotting was made using anti -glutathione - S - transferase (GST) antibodies, to detect the production of Pv -Def peptide (of ~ 4 KDa) linked to the GST tag (~ 26 KDa) of an approximate total molecular weight ~30 KDa. The Pv -Def peptide was successfully cleaved from the GST -tag. The Pv -Def induced growth inhibition of multi -drug resistant (MDR) bacterial culture Klebsiella spp , E. Coli and Staphylococcus aureus and fungal culture Aspergillus flavus and MDR Candida albicans.
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