Characterization and Sequencing of MT-Cox1 Gene in Khorasan Native Poultry

Abstract

The aim of this study was to investigate the nucleotide sequence of COX1 gene in mitochondrial genome of Khorasan native chicken and detect the possible mutations in the genome. For this purpose, after sampling and extracting DNA from the whole blood samples, the COX1 gene was amplified using specific primers and cloned in pTG19-T plasmid of the DH5α strain of Escherichia coli bacteria. Finally, the recombinant plasmids were extracted from positive colonies and sequenced. The results of nucleotide sequence indicated maximum of similarity with the same sequence in the Gallus gallus mitochondrial genome. Comparing these two sequences identified the presence of six nucleotide differences. However, only three point mutations led to the altering of dissimilar amino acids. Also, a protein BLAST query showed that the sequence had a similarity of 99% in 100% coverage to cytochrome C oxidase subunit 1. The protein BLAST query suggested the conserved domain cytochrome C, in a family which is part of a complex IV. This complex converts the oxygen into the water by electrons transfer from reduced cytochrome in a reaction associated with mitochondrial electron transport chain. The findings may be useful in identifying phenotypes in poultry.

Keywords: Cloning, Sequencing, MT-COX1, Khorasan native poultry, Gallus gallus