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Title 

Functional metagenome mining of soil for a novel gentamicin resistance gene

Authors 

Hyunjoo ImKyung Mo KimSang Heon LeeChoong-Min Ryu

Publisher 

The Korean Society for Applied Microbiology

Issue Date 

2016

Citation 

Journal of Microbiology and Biotechnology, vol. 26, no. 3, pp. 521-529

Keywords 

Antibiotic resistanceGentamicinMetagenomeRnlAShot-gun library

Abstract 

Extensive use of antibiotics over recent decades has led to bacterial resistance against antibiotics, including gentamicin, one of the most effective aminoglycosides. The emergence of resistance is problematic for hospitals, since gentamicin is an important broad-spectrum antibiotic for the control of bacterial pathogens in the clinic. Previous study to identify gentamicin resistance genes from environmental samples have been conducted using culturedependent screening methods. To overcome these limitations, we employed a metagenomebased culture-independent protocol to identify gentamicin resistance genes. Through functional screening of metagenome libraries derived from soil samples, a fosmid clone was selected as it conferred strong gentamicin resistance. To identify a specific functioning gene conferring gentamicin resistance from a selected fosmid clone (35-40 kb), a shot-gun library was constructed and four shot-gun clones (2-3 kb) were selected. Further characterization of these clones revealed that they contained sequences similar to that of the RNA ligase, T4 rnlA that is known as a toxin gene. The overexpression of the rnlA-like gene in Escherichia coli increased gentamicin resistance, indicating that this toxin gene modulates this trait. The results of our metagenome library analysis suggest that the rnlA-like gene may represent a new class of gentamicin resistance genes in pathogenic bacteria. In addition, we demonstrate that the soil metagenome can provide an important resource for the identification of antibiotic resistance genes, which are valuable molecular targets in efforts to overcome antibiotic resistance.

ISSN 

1017-7825

Link 

http://dx.doi.org/10.4014/jmb.1511.11053

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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