Journal of Applied Biosciences (J. Appl. Biosci.) [ISSN 1997 - 5902]
Volume 21: 1258 - 1264. Published September 7, 2009.
Sequence of specific mitochondrial 12S rRNA fragment of Egyptian buffalo as a reference for discrimination between buffalo, cattle, sheep and goat
*Hassan A. I. Ramadan and Eman R. Mahfouz
Department of Cell Biology, Genetic Engineering and Biotechnology Division, National Research Center, P.O. Box 12622, Dokki, Giza, Egypt.
*Corresponding author e-mail: hameguid@yahoo.comABSTRACT
Objective: Polymerase Chain Reaction (PCR) was conducted to amplify segment of specific mitochondrial 12S rRNA gene fragment in Egyptian water buffalo towards developing a tool for discriminating buffalo from cattle, sheep and goat.
Methodology and results: Amplification was performed using purified total genomic DNA of nine Egyptian buffalo blood samples. Direct sequence analysis showed complete conservation among all tested samples. The investigated buffalo sequences were deposited in GenBank database with the accession numbers (FJ828575-FJ828583). Multiple sequence alignment between the sequenced Egyptian buffalo and the homologous segments of buffalo, cattle, sheep and goat sequences available on GenBank database showed that the Egyptian buffaloes belong to one of two haplotypes, while the other one includes swamp buffalo. On the other hand definite single nucleotide polymorphisms (SNPs) were detected and were found to be specific for each of the four studied Bovidae species. In addition, other specific SNPs were detected that allow the discrimination between subfamily Bovinae, represented by water buffalo and cattle, and subfamily Caprinae represented by sheep and goat.
Conclusion and application of findings: Sequence examination and detection of specific SNPs referring to the specific Egyptian buffalo 12S rRNA gene fragment (accession numbers FJ828575-FJ828583) is an easy, accurate and highly specific method for the discrimination and identification of unknown sequences belonging to one of the four studied Bovidae species.
Key words: Sequence analysis, Bovidae, species identification, mitochondrial DNA, 12S rRNA gene.
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