EVALUATION OF SIX CAMEL BREEDS FOR HETEROZYGOCITY THROUGH RESTRICTION FRAGMENT LENGTH POLYMORPHISM

Abstract

In the camel tyrosinase gene, a restriction site provoked by the T variant was used in a special restriction fragment length polymorphism analysis (PCR-RFLP) for genotyping of animals from six different Pakistani camel breeds (Marecha, Dhatti, Larri, Kohi, Campbelpuri and Sakrai). For this purpose, four new primer pairs were designed for sequencing the coding region of exon 1 of the tyrosinase gene. PCR reactions were carried out in a total volume of 25 l using 100 ng genomic DNA to amplify a 474 bp fragment at 56°C. A SNP (T/C) at 200 bps was found and exploited with a Dde I restriction enzyme that resulted in three different genotypes i.e. TT, TC and CC in each studied camel breed. Significant differences in the genotype frequency between the breeds were recorded. The Sakrai breed showed a distinctly higher frequency of heterozygous animals compared to Marecha, Dhatti, Larri and Kohi breeds. Our new designed primers could be used for genotype screening of other camel breeds. However, for understanding the contribution of tyrosinase gene and its antagonist i.e. agouti in the coat colour production, complete sequence of the gene is imperative.

Keywords

• Camel
• genotyping
• RFLP
• tyrosinase