title: A novel screening system for secretion of heterologous proteins in Bacillus subtilis.
abstract: 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.
authors.affiliation: Department of Molecular Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands.
authors.forename: Hein
authors.initials: H
authors.lastname: Trip
authors.forename: Patricia J
authors.initials: PJ
authors.lastname: van der Veek
authors.forename: Ton C
authors.initials: TC
authors.lastname: Renniers
authors.forename: Rob
authors.initials: R
authors.lastname: Meima
authors.forename: Cees M
authors.initials: CM
authors.lastname: Sagt
authors.forename: Lisette
authors.initials: L
authors.lastname: Mohrmann
authors.forename: Oscar P
authors.initials: OP
authors.lastname: Kuipers
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D058947:Bacterial Secretion Systems; D005434:Flow Cytometry / Q000379:methods*; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism*
chemical_list: D058947:Bacterial Secretion Systems; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D000516:alpha-Amylases