Display of Multimeric Antimicrobial Peptides on the Escherichia coli Cell Surface and Its Application as Whole-Cell Antibiotics
Figure 1
Schematic representation of the construction of Lpp-OmpA-multimeric Buf IIIb-L fusion genes.
A monomeric Buf IIIb-L gene was dimerized by; (1) excision of the monomeric Buf IIIb-L insert by digestion with BbsI and FokI, (2) isolation of the fragment, (3) cloning into the original pMBT-B1 vector digested with BbsI, generating pMBT-B2. These steps were repeated for the construction of tandem multimers of the Buf IIIb-L gene, generating pMBT-Bn (n = number of Buf IIIb-L genes). The BbsI and FokI fragments of pMBT-Bn were cloned into pLpp-OmpA digested with BbsI, generating pLpp-OmpA-Bn. The NdeI and BamHI fragments of pLpp-OmpA-Bn were then ligated into the expression vector pET21c digested with the same enzymes, generating pLO-Bn (n = 0, 1, 2, and 3). The arrows, open triangles and closed triangles indicate BbsI cleavage sites and FokI cleavage sites, respectively.