Identification of Multiple Bacteriocins in Enterococcus spp. Using an Enterococcus-Specific Bacteriocin PCR Array
Abstract
:1. Introduction
2. Experimental Section
2.1. Bacterial Strains, Storage, and Growth Conditions
2.2. Activity against Listeria Monocytogenes
2.3. Primer Creation
2.4. PCR Detection of Enterococcal Bacteriocin Genes
3. Results
3.1. Primer Design
3.2. PCR Detection of Enterococcal Bacteriocin Genes by Real-Time PCR
3.3. Inhibition of L. monocytogenes 39-2 R0 (Wild-Type) and R1 (curvaticin FS47R)
Primer | Target Gene | Sequence (5′→3′) | Product Size (bp) | Included Genes | Homologous Genes |
---|---|---|---|---|---|
1 | Avicin A (avcA)—FJ851402.1 | 236 | avicin A precurson (avcA) immunity protein (avcI) divergicin-like bact (avcB) | sakacin X (sakX) | |
Forward | ACG CGA AAT GAA GAA TGT TG | ||||
Reverse | TTT CAT TTC CGC CAG AAA AC | ||||
2 | Columbicin A (colA)—EF033111.1 | 299 | columbicin A (colA) hypothetical protein (orfB) | bovicin A (bovA) enterocin AS-48 | |
Forward | TTT TTC TTG GGT TAT TTA CAG GAA | ||||
Reverse | ATG TGC AAT GGG CAA AAA CT | ||||
3 | Durancin Q (duqQ)—AB284369.1 | 384 | immunity protein (duqI) durancin Q (duqQ) inducing peptide (duqF) | durancin TW-49 (durM) | |
Forward | GCA CTG ATT CCG GCA CTA AT | ||||
Reverse | CGT AAC TCT AAT GGC GGG AAG | ||||
4 | Enterocin 96 (ent96)—FJ769024.1 | 291 | enterocin 96 | - | |
Forward | GTG GAG AGG ACG AAA GGA GA | ||||
Reverse | TTG ATT AGT GGA GAG GAC GGT TA | ||||
5 | Enterocin mr10A/mr10B (mr10AB) | 247 | enterocin mr10A (mr10A) enterocin mr10B (mr10B) | enterocin JSB (entJSB) enterocin NA (entNA) enterocin NB (entNB) enterocin L50A (entL50A) enterocin L50B (entL50B) enterocin 62-6A (ent626A) enterocin 62-6B (ent626B) enterocin RJ-11 | |
Forward | ATG GGA GCA ATC GCA AAA T | ||||
Reverse | CAT CCT TGT CCG ATA AAC TGC | ||||
6 | Enterocin C (entC)—FU862242.1 | 506 | enterocin C1 (entC1) enterocin C2 (entC2) enterocin C immunity (entCI) | enterocin 1081A enterocin 1071B | |
Forward | AGG TCC AGC TGC TTA TTG GA | ||||
Reverse | CCA TTA GAA TGA ATA CGC TAA AGA AA | ||||
7 | Enterocin SE-K4 (entSE-K4)—AB092692.1 | 608 | enterocin SE-K4 (entSE-K4) enterocin precursor (orf7) entSE-K4 homologue (orf8) entSE-K4 immunity (orf9) | bacteriocin II (D78257.1) enterocin TW-21 bacteriocin 31 | |
Forward | ATG TAG AAG CCG CCA CGT AT | ||||
Reverse | AAT CCC AAT CAT CCC ACA AA | ||||
8 | Enterocin EJ97(ej97a)—AJ490170.1 | 104 | enterocin ej97 | - | |
Forward | AAA GCG ATG ATT AAG AAG TTT CC | ||||
Reverse | TCC CAA GGA TAA CGA CCG TA | ||||
9 | Enterocin Wα/Wβ (enwAB)—AB600897.1 | 423 | enterocin W alpha (enwA) enterocin W beta (enw B) | - | |
Forward | GGG GTT GAA TTA TTG TAG AAA GGA | ||||
Reverse | AAC TAG CCT CTA CCG CCA CA | ||||
10 | Enterocin Q (entqA)—DQ832184.1 | 231 | enterocin Q (entqA) | - | |
Forward | ATC ACA AAG TGA GCC CCT GT | ||||
Reverse | TGG TAT CGC AAA ATG GAT GA | ||||
11 | Enterocin P (entP)—AF005726.1 | 431 | enterocin P (entP) enterocin P immunity (entQ) | - | |
Forward | TTC CCC GAA GAA TAC AAA TGA | ||||
Reverse | AAT TTC TGG GGT GGC TAA TG | ||||
12 | Enterocin A (entA)—AF240561.1 | 362 | enterocin A (entA) immunity protein (entI) | - | |
Forward | AAA ATA AAT GTA CGG TCG ATT GG | ||||
Reverse | CCA GCA GTT CTT CCA ATT TCA | ||||
13 | Enterocin B (entB)—U87997,1 | 257 | enterocin B (entB) | enterocin CRL35 | |
Forward | CAG AGT TCC CAA CTG TTT GCT | ||||
Reverse | AGC CCA TGC TAG TGG TCCT T | ||||
14 | Enterocin Xα/Xβ (enxAB)—AB430879.1 | 321 | enterocin X alpha (enxA) enteorcin X beta (enxB) | - | |
Forward | GGACAATTTATGGGTAAACAAGC | ||||
Reverse | TACGTCCACCATTCCAACCT | ||||
15 | Bacteriocin T8 (bacA)—AB178871.1 | 469 | bacteriocin precursor (bacA) hypothetical immunity protein (bacB) | hiracin JM79 Bac43 | |
Forward | TTGTCTAGCTGGCATCGGTA | ||||
Reverse | CCAATAGAAGCCCATCCTCT | ||||
16 | Mundticin KS (munA)—KC291253.1 | 285 | mundticin KS (munA) | mundticin L (munL) enterocin HF | |
Forward | AAA AGG GTG CAG TGT TGA TTG | ||||
Reverse | TCC ACT GAA ATC CAT GAA TGA |
Isolate | Species | entA | mr10AB | enxAB | bacA | entP | entB | munA |
---|---|---|---|---|---|---|---|---|
FS707 | En. durans | 99% | ||||||
BJ-12 | En. faecalis | 100% 2 | ||||||
BJ-13 | En. faecalis | 99% | ||||||
BJ-19 | En. faecalis | 100% | ||||||
BJ-27 | En. faecalis | 100% | 96% | |||||
326F | En. faecium | 100% | 92% | 99% | 99% | |||
FS56-1 | En. faecium | 100% | 97% | |||||
FS97-2 | En. faecium | 100% | 99% | 100% | ||||
JCP B-5 | En. faecium | 97% | 100% | 99% | ||||
JCP M-2 | En. faecium | 95% | 100% | 99% | ||||
JCP-9 | En. faecium | 98% | 100% | 99% | ||||
Milk12 | En. faecium | 100% | 99% | 100% | ||||
Milk5 | En. faecium | 100% | 99% | 100% | 99% | |||
NP-7 | En. faecium | 100% | ||||||
Pop4 | En. faecium | 100% | 96% | 100% | 99% | |||
THYME2 | En. faecium | 100% | 100% | |||||
THYME3 | En. faecium | 100% | ||||||
323F | En. hirae | 100% | 97% | |||||
323RL1 | En. hirae | 100% | 96% | 99% | ||||
341FA | En. hirae | 100% | 93% | 99% | ||||
FS92 | En. thailandicus | 100% | 97% | |||||
RP-1 | En. thailandicus | 100% |
4. Discussion
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Henning, C.; Gautam, D.; Muriana, P. Identification of Multiple Bacteriocins in Enterococcus spp. Using an Enterococcus-Specific Bacteriocin PCR Array. Microorganisms 2015, 3, 1-16. https://doi.org/10.3390/microorganisms3010001
Henning C, Gautam D, Muriana P. Identification of Multiple Bacteriocins in Enterococcus spp. Using an Enterococcus-Specific Bacteriocin PCR Array. Microorganisms. 2015; 3(1):1-16. https://doi.org/10.3390/microorganisms3010001
Chicago/Turabian StyleHenning, Chris, Dhiraj Gautam, and Peter Muriana. 2015. "Identification of Multiple Bacteriocins in Enterococcus spp. Using an Enterococcus-Specific Bacteriocin PCR Array" Microorganisms 3, no. 1: 1-16. https://doi.org/10.3390/microorganisms3010001