ARS | Publication request: Putative and unique gene sequence utilization for the design of species specific probes as modeled by Lactobacillus plantarum

Submitted to: Current Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: November 6, 2012
Publication Date: March 1, 2013
Repository URL: http://handle.nal.usda.gov/10113/55929
Citation: Perez Diaz, I.M. 2013. Putative and unique gene sequence utilization for the design of species specific probes as modeled by Lactobacillus plantarum. Current Microbiology. 66:266-270.

Interpretive Summary: The L. plantarum genome sequence was screened for unique genes, to be used as a tool for its identification in mixed cultures. The sequence of the unique gene was used to construct a labeled probe that would bind to the complimentary sequence when L. plantarum DNA is present on a membrane and generate a blue dot. If the labeled probe encounters DNA from other microbes, it will not bind or generate the blue dot.

Technical Abstract: The concept of utilizing putative and unique gene sequences for the design of species specific probes was tested. The abundance profile of assigned functions within the Lactobacillus plantarum genome was used for the identification of the putative and unique gene sequence, csh. The targeted gene (csh) was used as the template for PCR amplification and construction of a non-radioactive DIG labeled probe. The csh derived probe aided in the preliminary and rapid identification of L. plantarum from mixed cultures by colony hybridization. The method described here for the rapid identification of L. plantarum can also be applied for the rapid detection of other bacteria if a unique gene sequence can be identified from its complete genome sequence.