Recombinant Klebsiella pneumoniae Phosphoethanolamine--lipid A transferase MCR-8.1 (mcr-8), partial

Clone the gene fragment corresponding to amino acids 177-565 of the klebsiella pneumoniae phosphoethanolamine--lipid A transferase MCR-8.1 (mcr-8) into an expression vector suitable for E. coli expression, ensuring that the vector includes an N-terminal 6xHis-tag for purification purposes. Transform the recombinant expression vector carrying the mcr-8 gene fragment into competent E. coli cells. Select transformed cells by plating them on agar plates containing appropriate antibiotics specific to the selection marker of the expression vector. Cultivate the selected E. coli cells in a suitable growth medium, under conditions favorable for protein expression. Induce protein expression using an appropriate inducer, such as IPTG. Harvest the E. coli cells by centrifugation and proceed to cell lysis to release the cellular contents, including the expressed recombinant mcr-8 protein. Assess the purity of the recombinant mcr-8 protein using SDS-PAGE analysis, which exhibits a purity level of over 90%.