title: Bacillus subtilis as a tool for screening soil metagenomic libraries for antimicrobial activities.
abstract: Finding new antimicrobial activities by functional metagenomics has been shown to depend on the heterologous host used to express the foreign DNA. Therefore, efforts are devoted to developing new tools for constructing metagenomic libraries in shuttle vectors replicatable in phylogenetically distinct hosts. Here we evaluated the use of the Escherichia coli-Bacillus subtilis shuttle vector pHT01 to construct a forest-soil metagenomic library. This library was screened in both hosts for antimicrobial activities against four opportunistic bacteria: Proteus vulgaris, Bacillus cereus, Staphylococcus epidermidis, and Micrococcus luteus. A new antibacterial activity against B. cereus was found upon screening in B. subtilis. The new antimicrobial agent, sensitive to proteinase K, was not active when the corresponding DNA fragment was expressed in E. coli. Our results validate the use of pHT01 as a shuttle vector and B. subtilis as a host to isolate new activities by functional metagenomics.
authors.affiliation: Microbiology and Genomics Unit, Gembloux Agro-Bio Tech, University of Liège, B-5030 Gembloux, Belgium. Sophie.Biver@ulg.ac.be
authors.forename: Sophie
authors.initials: S
authors.lastname: Biver
authors.forename: Sébastien
authors.initials: S
authors.lastname: Steels
authors.forename: Daniel
authors.initials: D
authors.lastname: Portetelle
authors.forename: Micheline
authors.initials: M
authors.lastname: Vandenbol
mesh_terms: D000890:Anti-Infective Agents / Q000378:metabolism*; D001409:Bacillus cereus / Q000187:drug effects; D001412:Bacillus subtilis / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D015723:Gene Library; D005822:Genetic Vectors; D054892:Metagenome; D016982:Micrococcus luteus / Q000187:drug effects; D008969:Molecular Sequence Data; D011514:Proteus vulgaris / Q000187:drug effects; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D013212:Staphylococcus epidermidis / Q000187:drug effects
chemical_list: D000890:Anti-Infective Agents; D004269:DNA, Bacterial