Arivaradarajan, P;
Warburton, PJ;
Paramasamy, G;
Nair, SP;
Allan, E;
Mullany, P;
(2016)
Identification of an antibacterial protein by functional screening of a human oral metagenomic library.
FEMS Microbiology Letters
, 363
(1)
10.1093/femsle/fnv200.
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Abstract
Screening of a bacterial artificial chromosome (BAC) library containing metagenomic DNA from 22 human plaque and saliva allowed the isolation of four clones producing antimicrobial activity. Three 23 of these were pigmented and encoded homologues of glutamyl-tRNA reductase (GluTR), an enzyme 24 involved in the C5 pathway leading to tetrapyrole synthesis, and one clone had antibacterial activity 25 with no pigmentation. The latter contained a BAC with an insert of 15.6 kb. Initial attempts to 26 localise the gene(s) responsible for antimicrobial activity by sub-cloning into pUC-based vectors 27 failed. A new plasmid for toxic gene expression (pTGEX) was designed enabling localisation of the 28 antibacterial activity to a 4.7 kb HindIII fragment. Transposon mutagenesis localised the gene to an 29 open reading frame of 483 bp designated antibacterial protein1 (abp1). Abp1 was 94% identical to a 30 hypothetical protein of Neisseria subflava (accession number WP_004519448.1). An E. coli clone 31 expressing Abp1 exhibited antibacterial activity against Bacillus subtilis BS78H, Staphylococcus 32 epidermidis NCTC 11964 and B4268 , and Staphylococcus aureus NCTC 12493,ATCC 35696 and NCTC 33 11561 . However no antibacterial activity was observed against Pseudomonas aeruginosa ATCC 34 9027, Neisseria subflava ATCC A1078, Escherichia coli K12 JM109, and BL21(DE3) Fusobacterium 35 Deleted: 36 Deleted: (methicillin resistant, MRSA) 37 nucleatum ATCC 25586 and NCTC 11326, Prevotella intermedia ATCC 25611, Veillonella parvula 38 ATCC 10790 or Lactobacillus casei NCTC 6375.
| Type: | Article |
|---|---|
| Title: | Identification of an antibacterial protein by functional screening of a human oral metagenomic library |
| Location: | England |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1093/femsle/fnv200 |
| Publisher version: | http://dx.doi.org/10.1093/femsle/fnv200 |
| Language: | English |
| Additional information: | Copyright © FEMS 2015. All rights reserved. This is a pre-copyedited, author-produced PDF of an article accepted for publication in FEMS Microbiology Letters following peer review. The version of record, Arivaradarajan, P; Warburton, PJ; Paramasamy, G; Nair, SP; Allan, E; Mullany, P; (2016) Identification of an antibacterial protein by functional screening of a human oral metagenomic library. FEMS Microbiology Letters, 363 (1), is available online at: 10.1093/femsle/fnv200 |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Eastman Dental Institute UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Eastman Dental Institute > Microbial Diseases |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/1475657 |
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