SuPER PCR

A web forum for questions and answers (hopefully!) regarding the Suicide Polymerase Endonuclease Restriction (SuPER) methodology.

Thursday, December 04, 2008

Manuscript Using SuPER-PCR

The SuPER method was recently used to expose the presence of Pseudomonas species in the digestive system of fruit flies. The digestive system is predominantly composed of Enterobacteriaceae and these are detected routinely with bacterial 16S rRNA gene primers. Behar et al. (2008) describe the application of an Enterobacteriaceae-specific SuPER primer to digest away the 16S rRNA genes of these organisms. Standard bacterial PCR was then able to detect Pseudomonas. This was confirmed with specific cultivation of these organisms. A nice study and application of the SuPER method.

Gut bacterial communities in the Mediterranean fruit fly (Ceratitis capitata) and their impact on host longevity

A. Behar, B. Yuval and E. Jurkevitch.

Abstract

Fruit flies (Diptera: Tephritidae) harbor stable bacterial communities in their digestive system, composed mainly of members of the Enterobacteriaceae. However, the Enterobacteriaceae are not the sole community in this habitat. We examined the hypothesis that Pseudomonas spp. form a cryptic community in the gut of Ceratitis capitata, the Mediterranean fruit fly (‘medfly’). Suicide polymerase restriction PCR (SuPER PCR), a novel culture-independent technique, revealed that Pseudomonas spp. form minor, common and stable communities within the medfly's gut. These include P. aeruginosa, a known pathogen of arthropods. Experimental inoculations with high levels of P. aeruginosa reduced the medfly's longevity while inoculations with members of the Enterobacteriaceae extended the fly's life.

Accordingly, we suggest that in addition to their possible contribution to the fly's nitrogen and carbon metabolism, development and copulatory success (as shown in previous studies), the Enterobacteriaceae community within the medfly's gut may also have an indirect contribution to host fitness by preventing the establishment or proliferation of pathogenic bacteria.