title: Identification, sequencing and comparative analysis of pBp15.S plasmid from the newly described entomopathogen Bacillus pumilus 15.1.
abstract: The Bacillus pumilus 15.1 strain, a recently described entomopathogenic strain active against Ceratitis capitata, contains at least two extrachromosomal elements, pBp15.1S and pBp15.1B. Given that B. pumilus is not a typical entomopathogenic bacterium, the acquisition of this extrachromosomal DNA may explain why B. pumilus 15.1 is toxic to an insect. One of the plasmids present in the strain, the pBp15.1S plasmid, was sub-cloned, sequenced and analyzed using bioinformatics to identify any potential virulence factor. The pBp15.1S plasmid was found to be 7785 bp in size with a GC content of 35.7% and 11 putative ORFs. A replication module typical of a small rolling circle plasmid and a sensing and regulatory system specific for plasmids was found in pBp15.1S. Additionally, we demonstrated the existence of ssDNA in plasmid preparations suggesting that pBp15.1S replicates by the small rolling circle mechanism. A gene cluster present in plasmid pPZZ84 from a distantly isolated B. pumilus strain was also present in pBp15.1S. The plasmid copy number of pBp15.1S in exponentially growing B. pumilus cells was determined to be 33 copies per chromosome. After an extensive plasmid characterization, no known virulence factor was found so a search in the other extrachromosomal elements of the bacteria is needed.
authors.affiliation: Institute of Biotechnology, Campus Fuentenueva s/n, University of Granada, 18071 Granada, Spain. Electronic address: dianacg85@yahoo.com.
authors.forename: Diana C
authors.initials: DC
authors.lastname: Garcia-Ramon
authors.affiliation: Institute of Biotechnology, Campus Fuentenueva s/n, University of Granada, 18071 Granada, Spain. Electronic address: mjlunavas@correo.ugr.es.
authors.forename: Maria Jose
authors.initials: MJ
authors.lastname: Luque-Navas
authors.affiliation: International Center for Zoonoses (CIZ), Faculty of Veterinary Medicine and Zootechnic, Central University of Ecuador, PO Box.17-03-100, Quito, Ecuador. Electronic address: alf.molina7@gmail.com.
authors.forename: C Alfonso
authors.initials: CA
authors.lastname: Molina
authors.affiliation: Department of Computer Science and Artificial Intelligence, Campus Fuentenueva s/n, University of Granada, 18071 Granada, Spain. Electronic address: delval@decsai.ugr.es.
authors.forename: Coral
authors.initials: C
authors.lastname: Del Val
authors.affiliation: Institute of Biotechnology, Campus Fuentenueva s/n, University of Granada, 18071 Granada, Spain. Electronic address: aosuna@ugr.es.
authors.forename: Antonio
authors.initials: A
authors.lastname: Osuna
authors.affiliation: Institute of Biotechnology, Campus Fuentenueva s/n, University of Granada, 18071 Granada, Spain; Department of Biochemistry and Molecular Biology I, Campus Fuentenueva s/n, University of Granada, 18071 Granada, Spain. Electronic address: svt@ugr.es.
authors.forename: Susana
authors.initials: S
authors.lastname: Vilchez
mesh_terms: D001407:Bacillus / Q000235:genetics*; D001482:Base Composition / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded / Q000302:isolation & purification; D018628:Gene Dosage / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D022661:RNA, Untranslated / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D004277:DNA, Single-Stranded; D022661:RNA, Untranslated
keywords: Bacillus pumilus plasmid; RapA; RapAB; Rep; Small rolling circle plasmid; Virulence factors