Construction of bacterial artificial chromosome library from electrochemical microorganisms.
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.
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Jung Ho
JH
Biomedical Research Center, Korea Institute of Science and Technology, P. O. Box 131, Cheongryang, Seoul, Korea.
Kim
Man Su
MS
Cho
Hyuk
H
Chang
In Seop
IS
Lee
Jiyoung
J
Kim
Kyung Sik
KS
Kim
Byung Hong
BH
Park
Young In
YI
Han
Ye Sun
YS
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
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D038181:FMN Reductase; C023903:ferric citrate iron reductase