Schmitt Paulina, Mercado Luis, Díaz Mauricio, Guzmán Fanny, Arenas Gloria, Marshall Sergio H
Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2950, Valparaíso, Chile.
Peptides. 2008 Apr;29(4):512-9. doi: 10.1016/j.peptides.2007.12.012. Epub 2008 Jan 4.
CECdir-CECret is a novel non-toxic doublet 8.5 kDa peptide representing the natural coding sequence of the antimicrobial peptide Cecropin A from Drosophila melanogaster fused in-frame to its own inverted version. Expression of this cloned doublet peptide in Escherichia coli, yielded peptides that were mostly packaged into inclusion bodies. The new molecule was purified, solubilized and refolded, through a standard guanidine-based procedure. The recovered refolded peptides were then characterized by HPLC chromatography, MALDI-TOF-mass spectrometry and peptide sequencing, and finally evaluated for their antimicrobial potential. The novel doublet peptide CECdir-CECret, displays an enhanced in vitro antimicrobial activity and action spectrum in comparison to the monomer Cecropin A.
CECdir-CECret是一种新型无毒双联体8.5 kDa肽，它代表了来自黑腹果蝇的抗菌肽天蚕素A的天然编码序列，并与其自身的反向序列读框融合。这种克隆的双联体肽在大肠杆菌中表达，产生的肽大多被包装到包涵体中。通过基于胍的标准程序对新分子进行纯化、溶解和重折叠。然后通过高效液相色谱、基质辅助激光解吸电离飞行时间质谱和肽测序对回收的重折叠肽进行表征，最后评估其抗菌潜力。与单体天蚕素A相比，新型双联体肽CECdir-CECret在体外显示出增强的抗菌活性和作用谱。
