Polymorphic status and phylogenetic analysis of myostatin gene in pak-thoroughbred

Abstract

Myostatin is a protein translated by the the MSTN gene (also known as GDF8), is responsible for limiting muscle growth and strength. In thoroughbred horse (Equus Caballus), limited studies have been designed to examine the variants in the coding region of MSTN gene. However, no data is available regarding the single nucleotide polymorphisms (SNPs) of the MSTN about racing performance in thoroughbred horses of Pakistan. In this study blood samples of fifteen Pakistani thoroughbred horses were collected from Race Club Lahore and immediately transferred into the ice box. The DNA was extracted by using phenol-chloroform method. Primers were designed for the amplification of all exons of the MSTN gene. The amplified PCR products were precipitated and sequenced for the identification of SNPs. SNPs were identified by visualizing the peaks of sequenced data by using Chromas Software. Phylogenetic analysis of MSTN gene in Pak-thoroughbred with racing species and some breeds of horses like Marwari Indian breed, Sindhi breed, Kathlawari breed, Italian breed and Chines breed was done separately by using MEGA 6 software. The analysis of identified SNPs were carried out by software SNPator. The sequenced data with altered protein was published in GenBank with accession number MN604194. Results have shown a total of 3 single nucleotide polymorphisms through Blast with reference sequences. Two SNPs were found in exon 2 at position of 2406 (C/T) and 2408 (C/T) respectively. One SNP (T/C) was detected in exon 3 at the position of 4661. In conclusion, Pak-thoroughbred horse population has 3 polymorphisms in their coding region which can be used as a biological marker for athletic abilities in Pak- thoroughbred.