Development of a PCR-based assay for rapid detection of class IIa bacteriocin genes.
OBJECTIVE
We have developed a PCR-based assay using custom designed panel of primers which allows rapid detection of class IIa bacteriocin-coding genes. To demonstrate the applicability of the developed assay, the method was applied on 40 metagenomic DNA preparations isolated from native microbiota of Polish artisanal cheeses produced in the Tatra Mountains.


RESULTS
The developed assay was designed on the basis of a large scale alignment of class IIa bacteriocin-coding genes. A panel of seven primer pairs with confirmed ability to detect class IIa bacteriocin-coding sequences was obtained. The following study has revealed a superb bacteriocinogenic potential of all forty analysed cheese samples.


CONCLUSIONS
The majority of obtained sequences were lactic acid bacteria (LAB) related, although some sequences showed significant similarity to bacteriocin-coding sequences present in non-LAB bacteriocin producers. The results suggest that several potentially new bacteriocin-coding sequences were found.


CONCLUSIONS
The developed assay can be extremely helpful in establishing whether isolates from the environment of interest have a potential of synthesizing antilisterial class IIa bacteriocins. Application of the approach may represent a useful tool contributing to ecological studies looking for valuable probiotic, bacteriocinogenic microbiota developing in foods.
Department of Biotechnology and Food Microbiology, Poznań University of Life Sciences, Wojska Polskiego 48, Poznań, Poland. wieckom@up.poznan.pl
Michał
M
Więckowicz
Marcin
M
Schmidt
Anna
A
Sip
Włodzimierz
W
Grajek
D001430:Bacteriocins / Q000235:genetics / Q000302:isolation & purification*; D002611:Cheese / Q000382:microbiology*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D005516:Food Microbiology; D005798:Genes, Bacterial; D007777:Lactobacillaceae / Q000235:genetics* / Q000378:metabolism; D054892:Metagenome; D011044:Poland; D016133:Polymerase Chain Reaction / Q000379:methods*; D019936:Probiotics
D001430:Bacteriocins; D017931:DNA Primers; D004269:DNA, Bacterial