Development of a DNA marker by minisatellite associated sequence amplification (MASA) from the endangered Indian rhino (Rhinoceros unicornis)

Abstract

Rhinoceroses are highly endangered species and their protection warrants immediate remedial measures. Development of DNA markers is envisaged to complement global efforts of the conservation of these extant animals. Minisatellite associated sequence amplification (MASA) of DNA from Indian rhinoceros (Rhinoceros unicornis) and three sub-species of South African black rhinoceros (Diceros bicornis) was carried out using a primer based on consensus sequence of the minisatellite repeat locus 33.15. Several bands in the range of 3.0 kilobases (kb) to 650 base pairs (bp) were identified that were useful for successful differentiation of R. unicornis from D. bicornis. Of these fragments, a 688 bp one, unique to R. unicornis was cloned and sequenced (Accession No. AF-296689). The band patterns uncovered by MASA and the species-specific hybridisation of pSG5 may be utilised as a tool for differentiating the R. unicornis genome from that of D. bicornis. This approach may also be adopted for the development of DNA-based genetic marker(s) useful for identification of other endangered species.