A vector system for ABC transporter-mediated secretion and purification of recombinant proteins in Pseudomonas species.

ABSTRACT: Pseudomonas fluorescens is an efficient platform for recombinant protein production. P. fluorescens has an ABC transporter secreting endogenous thermostable lipase (TliA) and protease, which can be exploited to transport recombinant proteins across the cell membrane. In this study, the expression vector pDART was constructed by inserting tliDEF, genes encoding the ABC transporter, along with the construct of the lipase ABC transporter recognition domain (LARD), into pDSK519, a widely used shuttle vector. When the gene for the target protein was inserted into the vector, the C-terminally fused LARD allowed it to be secreted through the ABC transporter into the extracellular medium. After secretion of the fused target protein, the LARD containing a hydrophobic C terminus enabled its purifica

SUBMITTER: Ryu J 

PROVIDER: S-EPMC4325144 | biostudies-literature | 2015 Mar

REPOSITORIES: biostudies-literature