Part:BBa_K2622043:Experience

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Applications of BBa_K2622043

Amplified gene sequence of BamC was ligated into a plasmid pCDFDuet. pelB signal sequence of BamC directs protein into periplasm. BamC can be purified in subcomplex with BamD and BamE using histidine tag only on BamE. To purify BamC, E. coli cells we induced with 0,1mM IPTG, grown in 37*C for 3 hours, then pelleted and sonicated, membrane fraction was isolated by centrifugation and was further purified with Ni-NTA and gelfiltration. BamC was used in a folding assay, which showed that BamB and BamCDE lipoproteins can assemble BamA into the membrane with an efficiency higher than 50% in time range of only two hours.

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