Open AccessA novel screening system for secretion of heterologous proteins in Bacillus subtilis
Author(s) -
Trip Hein,
van der Veek Patricia J.,
Renniers Ton C.,
Meima Rob,
Sagt Cees M.,
Mohrmann Lisette,
Kuipers Oscar P.
Publication year - 2011
Publication title -
microbial biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.287
H-Index - 74
ISSN - 1751-7915
DOI - 10.1111/j.1751-7915.2011.00270.x
Subject(s) - heterologous , bacillus subtilis , green fluorescent protein , secretion , secretory protein , biology , gene , reporter gene , heterologous expression , microbiology and biotechnology , strain (injury) , biochemistry , bacteria , recombinant dna , gene expression , genetics , anatomy
Summary High‐level production of secretory proteins in Bacillus subtilis leads to a stress response involving the two‐component system CssRS and its target genes htrA and htrB . Here, we used this sensing system in a reporter strain in which gfp is under control of P htrA , the secretion stress responsive promoter of htrA . Overexpression of heterologous secretory proteins in this strain results in green fluorescent cells, which can be separated from non‐secreting, low fluorescent cells using a fluorescence‐activated cell sorter (FACS). Using this principle, genomic libraries of uncharacterized prokaryotic organisms, expressed in the reporter strain, can be screened for genes encoding secretory proteins.