The hawksbill turtle Eretemochelys imbricata circumtropical is a species associated with coral reefs and other coastal habitats of the Atlantic Ocean, in the Colombian Caribbean. During their life cycle, the turtles migrate thousands of miles, traveling regularly to the high seas and territorial waters of different countries.
Due to human pressures, bycatch in fishing gear industry, demand for the shell and the plaston for its handicrafts, the marketing of eggs, meat, oil and habitat degradation have made this species is critically endangered with extinction (IUCN, 1994). This condition is very important to identify any level of this chain to the hawksbill turtle, and thus contribute to their conservation. In this study we used the COI gene as a molecular tag for identification using RFLP and sequencing. In the short term, this methodology will allow combat the illegal trade in specimens of E. imbricata and the use of derivatives.
Materials and Methods
We used blood samples from 15 juvenile hawksbill turtle from the island of St. Martin de Pajarales (Colombian Caribbean) (Fig. 2). Total DNA was extracted and amplified by PCR the mitochondrial gene cytochrome c oxidase subunit I (COI). The amplified fragments were analyzed by RFLP using the restriction enzyme AluI. Then compared in silico with the COI gene of Chelonia mydas and other species of freshwater turtles (GenBank). Were obtained COI gene sequences for each individual studied and analyzed by BLAST to identify genus and species.
DNA was obtained in good quality and concentration (50 ng/ul), being optimal for use in PCR reactions. Were amplified 12 fragments between 700-835 bp (Figure 4), corresponding to the COI mitochondrial gene.
The product obtained by PCR of the COI gene was cut with restriction enzyme AluI produced three bands of 352, 308 and 172 bp. This pattern was compared with that obtained bioinformatics in silico (NEBCutter v.o2) (Figure 5), showing...