wrapping-it-up…

To this day we have about 80% of our work done.  We are still working on sequencing our clones.  After that, it’s all about bioinformatics… in order to interpret our results.  I hope that we can have the sequencing results in order to include them in my poster for the BioMinds Symposium!

 

Search for antibiotic resistance genes from Metagenomic Libraries Derived from Microbial Mats at the Cabo Rojo Salterns

Casanova Angel,^1* Robert Rabelo,¹ Lilliam Casillas,¹  Karen Cloud-Hansen², Heather K. Green² and Carlos Rios-Velazquez ³

¹University of PR-Humacao, ²University of Wisconsin-Madison, ³University of Puerto Rico-Mayaguez

The Cabo Rojo Salterns located at the Southwestern region of Puerto Rico posses a series of salt ponds surrounded by microbial mats where microorganisms are subjected to extreme conditions. According to prior phylogenetic studies, most microorganisms in the mats belong to novel unidentified groups which make the mats excellent locations for the construction of metagenomic libraries.  Direct cloning of environmental DNA from the mats might have great potential for identifying new genes and gene products for biotechnological purposes particularly those conferring resistance to antibiotics. To determine if novel antibiotic resistance genes can be found from the mats we constructed a metagenomic library from the mats of the Candelaria lagoon. Genomic DNA was extracted from the Candelaria microbial mats using an indirect method, and a fosmid library of approximately 33,000 clones was generated in Escherichia coli as a host. The clones in the fosmid library carried inserts of more than 20Kbp. The library was then screened for antibiotic resistance genes using agar plates supplemented with Carbenicillin and Gentamycin. From the initial screening, seven positive clones were chosen, tested for the presence of an insert, and reconfirmed for the antibiotic resistance.  To date the preliminary sequencing analysis conducted revealed genes of unrelated functions within the 20Kbp insert. Sequencing efforts are currently underway to determine the gene responsible for this activity and further determine if novel mechanisms for antibiotic resistance can be reported from these clones isolated from the Cabo Rojo salterns microbial mats.

~ by Angel on March 2, 2009.