title: Functional screening of a metagenomic library reveals operons responsible for enhanced intestinal colonization by gut commensal microbes.
abstract: Evidence suggests that gut microbes colonize the mammalian intestine through propagation as an adhesive microbial community. A bacterial artificial chromosome (BAC) library of murine bowel microbiota DNA in the surrogate host Escherichia coli DH10B was screened for enhanced adherence capability. Two out of 5,472 DH10B clones, 10G6 and 25G1, exhibited enhanced capabilities to adhere to inanimate surfaces in functional screens. DNA segments inserted into the 10G6 and 25G1 clones were 52 and 41 kb and included 47 and 41 protein-coding open reading frames (ORFs), respectively. DNA sequence alignments, tetranucleotide frequency, and codon usage analysis strongly suggest that these two DNA fragments are derived from species belonging to the genus Bacteroides. Consistent with this finding, a large portion of the predicted gene products were highly homologous to those of Bacteroides spp. Transposon mutagenesis and subsequent experiments that involved heterologous expression identified two operons associated with enhanced adherence. E. coli strains transformed with the 10a or 25b operon adhered to the surface of intestinal epithelium and colonized the mouse intestine more vigorously than did the control strain. This study has revealed the genetic determinants of unknown commensals (probably resembling Bacteroides species) that enhance the ability of the bacteria to colonize the murine bowel.
authors.affiliation: Department of Microbiology, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors.forename: Mi Young
authors.initials: MY
authors.lastname: Yoon
authors.forename: Kang-Mu
authors.initials: KM
authors.lastname: Lee
authors.forename: Yujin
authors.initials: Y
authors.lastname: Yoon
authors.forename: Junhyeok
authors.initials: J
authors.lastname: Go
authors.forename: Yongjin
authors.initials: Y
authors.lastname: Park
authors.forename: Yong-Joon
authors.initials: YJ
authors.lastname: Cho
authors.forename: Gerald W
authors.initials: GW
authors.lastname: Tannock
authors.forename: Sang Sun
authors.initials: SS
authors.lastname: Yoon
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D018441:Biofilms / Q000254:growth & development*; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D003062:Codon / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000502:physiology; D015723:Gene Library; D007420:Intestine, Large / Q000382:microbiology*; D054892:Metagenome / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016366:Open Reading Frames / Q000235:genetics; D009876:Operon / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D003062:Codon; D017931:DNA Primers