Composite

Part:BBa_K2622039:Experience

Designed by: Auksė Gaižauskaitė   Group: iGEM18_Vilnius-Lithuania   (2018-10-10)


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Applications of BBa_K2622039

Amplified gene sequence of BamA was ligated into pET28b plasmid. Protein expression was induced by 0,1mM IPTG and overexpressed in cytoplams in a form of inclusion bodies. Bacteria cells were lysed by sonication, inclusion bodies were isolated by centrifugation and dissolved in 8M urea. This yielded high purity of BamA and therefore did not need any further purification. BamA was used in a folding assay, which have shown that BamB and BamCDE lipoproteins can assemble BamA into the membrane with an efficiency higher than 50% in time range of only two hours.

User Reviews

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