Species identification by means of the cytochrome b gene

Abstract

Species identification was carried out by nucleotide sequence analysis of the cytochrome b (cytb) gene. The aim of the study was to identify biological specimens from diverse vertebrate animals by extracting and amplifying DNA from 44 different animal species covering the 5 major vertebrate groups (i.e. mammals, birds, reptiles, amphibians and fishes). The sequences derived were used to identify the biological origin of the samples by aligning to cytb gene sequence entries in nucleotide databases using the program BLAST. All sequences were submitted to the GenBank including new species which were not observed in the databases. The applicability of this method to the forensic field is demonstrated by simulated casework conditions where different types of samples including problematic specimens such as hair, bone samples, bristles and feathers were investigated to identify the species.

Extração de DNA e avaliação da composição espécie-específica de queijos

“…O método do fenol:clorofórmio apenas precipita o DNA, mantendo os demais inibidores. (Partis et al, 2000;Parson et al, 2000;Verkaar et al, 2002;Bellis et al, 2003;Girish et al, 2005).…”

“…A escolha de fragmentos do citocromo b mitocondrial para amplificação de DNA foi relevante, uma vez que este é mais abundante que o genômico, o que aumenta a sensibilidade da técnica, levando-se em consideração que as amostras de DNA são obtidas de poucas células somáticas provenientes da descamação natural do úbere de vacas saudáveis (Parson et al, 2000;Partis et al, 2000;Herman, 2001;Girish et al, 2005;Arslan et al, 2006).…”

A Pioneer Case Study on Identification of Infant Rhinoceros Horn

“…In sequence matching approach, a measure of the similarity between sequences is calculated and the most similar species is attributed to the sample [42,43] A 187 bp consensus sequence of Cyt b was obtained from confiscated sample and a sequence similarity search performed in GenBank provided the highest similarity (100%) with Rhinoceros unicornis (NCBI Accession JN935370) i.e. an Asiatic one-horned Rhinoceros, whereas, it had 87% sequence similarity with Rhinoceros sondaicus (NCBI Accession: AJ245725) followed by other Rhinoceros species/subspecies with a decreasing sequence similarity (Table 1).…”

“…Appropriate Genetic DNA markers are those which are rather conservative within the species but discriminative between the species [44]. Cyt b shows limited variability within and much greater variation between species [45] thus ideal for species identification [42]. The phylogenetic tree is used to discern the evolutionary relationships between the suspected sample and reference sequences from each possible species [5,46,47].…”

Identificação de Espécies de Animais Silvestres por Meio de Sequências Mitocondriais Utilizados no Combate aos Crimes Contra a Fauna – Mato Grosso – Brasil

“…(F5'-TAAA-CTTCAGGTGACCAAAAAATCA-3') 8 Diversas regiões do DNA mitocondrial podem ser utilizadas na identificação de espécies animais, entre elas, o gene Cytocromo b (cytb), que tem sido uma escolha tradicional para este fim desde sua descoberta 22,23 . Na literatura, podem ser encontrados inúmeros exemplos do uso desta região na identificação de vertebrados para fins forenses, incluindo mamíferos 24 , peixes 25 ; répteis 26 , e aves 27,28 , inclusive seus ovos [29][30][31] .…”