RpoB-PCR amplified gene and temporal temperature gradient gel electrophoresis: a rapid tool to analyse bacterial strains representative of cold-smoked salmon microflora

Type Article
Date 2004-02
Language English
Author(s) Giacomazzi Sophie1, Leroi Francoise1, L'Henaff Cecile1, 2, Joffraud Jean-Jacques1
Affiliation(s) 1 : IFREMER, Lab Genie Alimentaire, F-44311 Nantes 3, France.
2 : ENITIAA, Nantes, France.
Source Letters in Applied Microbiology (0266-8254) (Blackwell science), 2004-02 , Vol. 38 , N. 2 , P. 130-134
DOI 10.1111/j.1472-765X.2003.01463.x
WOS© Times Cited 12
Keyword(s) TTGE, RpoB, Nested PCR, Cold smoked salmon flora
Abstract Aims: To evaluate rpoB gene as a biomarker of microbial biodiversity associated to cold-smoked salmon by a novel nested-polymerase chain reaction/temporal temperature gradient gel electrophoresis (PCR/TTGE) technique applied on pure cultures of reference strains.

Methods and Results: DNA obtained from pure cultures of reference strains was used in a succession of a first PCR amplification of rpoB fragment with degenerated nonclamped primers and a nested-PCR with nondegenerated clamped primers. PCR products were then applied on a TTGE gel in order to analyse strains profile. High quantity of nested-PCR products were obtained for each tested strain and TTGE profiles showed a good separation between the different reference bacteria and an easy way to associate one band to one species.

Conclusion: The nested-PCR/TTGE technique used in this study is a promising way of investigating bacterial community structure of cold-smoked salmon or other food matrix.

Significance and Impact of the Study: Because of its single copy state leading to single band profiles in TTGE, rpoB constitute a good potential molecular marker for further development of cold-smoked salmon biodiversity analysis.
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Giacomazzi Sophie, Leroi Francoise, L'Henaff Cecile, Joffraud Jean-Jacques (2004). RpoB-PCR amplified gene and temporal temperature gradient gel electrophoresis: a rapid tool to analyse bacterial strains representative of cold-smoked salmon microflora. Letters in Applied Microbiology, 38(2), 130-134. Publisher's official version : https://doi.org/10.1111/j.1472-765X.2003.01463.x , Open Access version : https://archimer.ifremer.fr/doc/00000/579/