title: Construction of bacterial artificial chromosome library from electrochemical microorganisms.
abstract: A microbial fuel cell is a device that directly converts metabolic energy into electricity, using electrochemical technology. The analysis of large genome fragments recovered directly from microbial communities represents one promising approach to characterizing uncultivated electrochemical microorganisms. To further assess the utility of this approach, we constructed large-insert (140 kb) bacterial artificial chromosome (BAC) libraries from the genomic DNA of a microbial fuel cell, which had been operated for three weeks using acetate media. We screened the expression of several ferric reductase activities in the Escherichia coli host, in order to determine the extent of heterologous expression of metal-ion-reducing enzymes in the library. Phylogenetic analysis of 16S rRNA gene sequences recovered from the BAC libraries indicates that they contain DNA from a wide diversity of microbial organisms. The constructed bacterial library proved a powerful tool for exploring metal-ion reductase activities, providing information on the electron transport pathway of electrochemical microbial (ECM) organisms.
authors.affiliation: Biomedical Research Center, Korea Institute of Science and Technology, P. O. Box 131, Cheongryang, Seoul, Korea.
authors.forename: Jung Ho
authors.initials: JH
authors.lastname: Back
authors.forename: Man Su
authors.initials: MS
authors.lastname: Kim
authors.forename: Hyuk
authors.initials: H
authors.lastname: Cho
authors.forename: In Seop
authors.initials: IS
authors.lastname: Chang
authors.forename: Jiyoung
authors.initials: J
authors.lastname: Lee
authors.forename: Kyung Sik
authors.initials: KS
authors.lastname: Kim
authors.forename: Byung Hong
authors.initials: BH
authors.lastname: Kim
authors.forename: Young In
authors.initials: YI
authors.lastname: Park
authors.forename: Ye Sun
authors.initials: YS
authors.lastname: Han
mesh_terms: D001419:Bacteria / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D022202:Chromosomes, Artificial, Bacterial; D002876:Chromosomes, Bacterial / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D038181:FMN Reductase / Q000235:genetics / Q000378:metabolism; D020459:Genes, rRNA; D016680:Genome, Bacterial; D015698:Genomic Library; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D038181:FMN Reductase; C023903:ferric citrate iron reductase