Open AccessConstruction of pMEKm12, an expression vector for protein production in Pseudomonas syringae
Author(s) -
Lu ShiEn,
ScholzSchroeder Brenda K.,
Gross Dennis C.
Publication year - 2002
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2002.tb11169.x
Subject(s) - pseudomonas syringae , biology , nonribosomal peptide , pseudomonas putida , maltose binding protein , fusion protein , expression vector , mutant , pseudomonas , gene , heterologous expression , escherichia coli , shuttle vector , vector (molecular biology) , microbiology and biotechnology , biochemistry , genetics , bacteria , recombinant dna , biosynthesis
Characterization of the biological roles of proteins is essential for functional genomics of pseudomonads. Heterologous proteins overproduced in Escherichia coli frequently fail to exhibit biological function. To circumvent this problem, vector pMEKm12 was constructed and used to overexpress proteins in Pseudomonas . The vector contains the pRO1600 replication origin, the maltose‐binding protein (MBP) fusion system, and an inducible tac promoter. The pMEKm12 was successfully used to overexpress the syringomycin synthetase SyrB1 protein fused to MBP in Pseudomonas syringae pv. syringae . Furthermore, expression of the MBP‐SyrB1 protein in the syrB1 mutant BR132A1 resulted in the restoration of syringomycin production. This vector will facilitate confirmation of the biochemical roles of nonribosomal peptide synthetase genes in Pseudomonas syringae , and studies of gene function from a wide spectrum of pseudomonads.