Cloning of mcr-1 and mcr-2 genes in Escherichia coli strain DH5-α to evaluate colistin resistance, protein pattern changes and plasmid stability
Abstract
Plasmid genes mcr-1 and mcr-2 are among the genes resistant to the antibiotic colistin, but Escherichia coli DH5-α lacks these genes and is therefore sensitive to the antibiotic colistin. In this study, the transformation of plasmid to E. coli DH5-α strain was performed using the transformation operation and the changes caused by antibiotic resistance or sensitivity, changes in structural and functional peptides and the stability of plasmids in the transformed strain were investigated.
Methods: The plasmids containing genes mcr-1 and mcr-2 were extracted from E. coli kp81 and kp37, respectively, and were transformed to E. coli DH5-α. To study their peptides using mass spectrometry, adsorption technique and laser ionization with MALDI-TOF MS matrix were performed.
Results: The results of transformation and proteomic results showed that antibiotic resistance to colistin was created by receiving and expressing colistin-resistant genes and changes in the peptides of the treated strain.