Genetic variability in exon 9 of PEPCK-M gene in different strains of chickens using PCR-RFLP method

Abstract

Phosphoenolpyruvate carboxykinase (PEPCK) catalyzes the first step of the gluconeogenesis cycle. There are two isozymes forms of this enzyme, cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) and mitochondrial phosphoenolpyruvate carboxykinase (PEPCK-M) which present in mitochondria and cytoplasm Current research has been conducted to identify the allelic polymorphism in the PEPCK-M gene in breeder hens of native fowls and commercial broiler and layer chickens. Blood samples were collected randomly from 150 birds of three strains and DNA was extracted using modified salting out method. A fragment of 401 bp in length was amplified from exon 9 of PEPCK-M gene. For genotyping of each sample the PCR products were digested by AccI restriction enzyme. The frequency of AccI- and AccI+ allele was estimated at 0.73 and 0.27 in native fowls population, 0.6 and 0.4 in broiler and 0.85 and 0.15 in layer lines, respectively. Two genotypes of AccI -/- and AccI -/+ were observed with the frequency of 0.46 and 0.54 in native fowls, 0.20 and 0.80 in broiler line and 0.70 and 0.30 in commercial layer line, respectively. The comparison of allelic frequency showed no statistical differences between native fowls population with broiler and layer lines, but this results indicated significantly differences between broiler and layer lines (P<0.05).The comparison of genotypic frequency showed that there were significant differences between three populations (P<0.05). No any AccI +/+genotype was detected in the genotyped samples. The difference in genotypic distribution may be as a consequence of different selection strategies used in these populations.

Keywords

• polymorphism
• posphoenolpyruvatecarboxykinase
• native fowls
• broiler
• layer