Two universal primer sets for species identification among vertebrates
Tóm tắt
The aim of this study was to develop a simple method using universal primers for species identification based on direct PCR sequencing. Two primer sets were designed based on the conserved regions of the 12S and 16S rRNA loci detected by the comprehensive sequence comparison among 30 mammalian whole mitochondrial genomes. In humans, the expected sizes of PCR products of the 12S and 16S rRNAs were 215 and 244 bp, respectively. Both primer sets successfully amplified the expected PCR products from various kinds of vertebrates including mammals, birds, reptiles, amphibians, and fish, and the sequenced segments contained sufficient nucleotide differences to identify each animal species. A case example of the identification of a piece of buried bone of unknown species is presented, and the species was identified as a pig by this method.
Tài liệu tham khảo
Zehner R, Zimmermann S, Mebs D (1998) RFLP and sequence analysis of the cytochrome b gene of selected animals and man: methodology and forensic application. Int J Legal Med 111:323–327
Bravi CM, Liron JP, Mirol PM, Ripoli MV, Peral-Garcia P, Giovambattista G (2004) A simple method for domestic animal identification in Argentina using PCR-RFLP analysis of cytochrome b gene. Leg Med (Tokyo) 6:246–251
Kocher TD, Thomas WK, Meyer A, Edwards SV, Pääbo S, Villablanca FX, Wilson AC (1989) Dynamics of mitochondrial DNA evolution in animals: amplification and sequencing with conserved primers. Proc Natl Acad Sci USA 86:6196–6200
Parson W, Pegoraro K, Niederstatter H, Föger M, Steinlechner M (2000) Species identification by means of the cytochrome b gene. Int J Legal Med 114:23–28
Hsieh HM, Chiang HL, Tsai LC, Lai SY, Huang NE, Linacre A, Lee JC (2001) Cytochrome b gene for species identification of the conservation animals. Forensic Sci Int 122:7–18
Branicki W, Kupiec T, Pawlowski R (2003) Validation of cytochrome b sequence analysis as a method of species identification. J Forensic Sci 48:83–87
Bellis C, Ashton KJ, Freney L, Blair B, Griffiths LR (2003) A molecular genetic approach for forensic animal species identification. Forensic Sci Int 134:99–108
Balitzki-Korte B, Anslinger K, Bartsch C, Rolf B (2005) Species identification by means of pyrosequencing the mitochondrial 12S rRNA gene. Int J Legal Med 119:291–294
Bataille M, Crainic K, Leterreux M, Durigon M, de Mazancourt P (1999) Multiplex amplification of mitochondrial DNA for human and species identification in forensic evaluation. Forensic Sci Int 99:165–170
Dalmasso A, Fontanella E, Piatti P, Civera T, Rosati S, Bottero MT (2004) A multiplex PCR assay for the identification of animal species in feedstuffs. Mol Cell Probes 18:81–87
Thompson JD, Gibson TJ, Higgins DG (1994) ClustalW: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680
Nei M (1987) Molecular evolutionary genetics. Columbia University Press, New York, pp 254–286
Kimura M (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Springer MS, Douzery E (1996) Secondary structure and patterns of evolution among mammalian mitochondrial 12S rRNA molecules. J Mol Evol 43:357–373
Burk A, Douzery EJP, Springer MS (2002) The secondary structure of mammalian mitochondrial 16S rRNA molecules: refinements based on a comparative phylogenetic approach. J Mamm Evol 9:225–252