Part:BBa_K5302015

pBBR-AIDA-mCherry

This work is derived from pBBR1MCS-2 and mCherry-middle, and it has undergone codon optimization. This composite part combines AIDA(approximately 63.8kda) and mCherry(26.7kda), and we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express mCherry as a fluorescence labeling.The plasmid uses lac promotor and has kanamycin resistence. AIDA-I (Adhesin involved in diffuse adherence-I) is a self-transporter from enteropathogenic Escherichia coli (EPEC), which displays the target protein on the bacterial surface through a self-transport mechanism. It contains several key domains: an N-terminal signal peptide, a passenger domain (acting outside the cell), a connecting domain and a transport domain, which is predicted to form a β barrel structure and integrate into the outer membrane, forming channels for transport of the passenger domain. The transport mechanism of AIDA-I can be exploited to display recombinant proteins on the surface of E. coli.

Figure 1. structure of AIDA, from genbank

Figure 2. Colony PCR results of pBBR1MCS-AIDA-mCherry

Sequence and Features

Assembly Compatibility:

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