FIG. 1.
Strategies for constructing protein expression and secretion strains of T. kodakarensis. (A) Amino acid sequence of the putative secretion signal in the N-terminal region of TK1675. Basic residues following the initial Met residue are indicated with + symbols, and the hydrophobic region and the cleavage signal Ala-Ser-Ala are underlined. (B) Strategy for constructing the gene cassette for overexpression and secretion of ChiAΔ4. (C) Strategies for integrating the ChiAΔ4 and PyrF expression/secretion gene cassettes into the T. kodakarensis KUW1 genome with pSecChiA and pSecPyrF, respectively. Tgdh represents the 3′-flanking region of the glutamate dehydrogenase gene (TK1431), inserted to promote transcription termination.