العزلة والتعرف والتوصيف والاستنساخ والإفراط في التعبير عن جينات النباتات ذات الأنشطة المضادة للميكروبات

Abstract

(MzDef) peptide from Egyptian Zea mays L. and evaluate the MzDef antibacterial and antifungal activities against some bacterial species and some phytopathogenic fungi as well as against some types of cancer cells.Two degenerate primers were designed according to the region of putative plant defensin complete coding sequence available in Genbank database were used to amplify the putative defensin DNA coding sequence. The amplified DNA band with the expected defensin size was excised from the agarose gel. The purified defensin PCR DNA product was set for sequencing. Then the defensin coding sequence was sub cloned in prokaryotic expression pGex-4t1vector. The resulted recombinant pGex-4t1 expression vector was transformed in Escherichia coli bacterial strains BL21 (DE3) pLysS + RIL. Defensin DNA coding sequence was inserted into frame to be fused with GST-tagged gene to facilitate afterward purification. The total proteins were extracted from induced Bl21 Escherichia coli after its transformation with recombinant pGex-4t1. The extracted total protein was separated by agarose gel. The presence of the expressed definsin-GST fusion protein was immuno detected with anti-GST antibodies.