FIG. 1.
(A) Structure of secretion vector pMM1525 encoding the signal peptide of the B. megaterium extracellular esterase LipA (SPlipA) and carrying a multiple cloning site. The amino acid sequence of the open reading frame (orf) and the cleavage site AXA of the signal peptidase type I are indicated. (B) Expression plasmids for secretion of tagged proteins of interest via the LipA signal peptide. All plasmids are based on the shuttle vector pMM1520 (1, 2). All expression plasmids shown allow parallel cloning of genes of interest into the identical multiple cloning sites, from BglII (marked in bold) to NgoMIV. (C) The elements for inducible gene expression in B. megaterium are the xylose-inducible promoter (PxylA) and the gene for the xylose repressor (xylR). The elements for plasmid replication in Bacillus sp. are the origin of plasmid replication (ori), a gene essential for plasmid replication (repU), and the tet resistance gene. The elements for plasmid replication in E. coli are the origin of replication, colE1, and the ampicillin resistance gene bla.