Detection of polymorphism in booroola gene and growth differentiation factor 9 in Lori sheep breed

Shahram Nanekarani,Majid Goodarzi,Salman Torabi,Saber Khederzadeh,Nasir Landy

doi:10.4314/tjpr.v15i8.2

Abstract

Purpose: This study was carried out for detection of possible polymorphisms in Booroola gene (FecB) and growth differentiation factor 9 [GDF9] in Lori sheep breed. Methods: Blood samples were taken from the jugular vein of the sheep and their DNA content extracted using modified salting-out method. The quantity and quality of extracted DNA was examined by absorption spectrophotometry and gel electrophoresis. Using two pairs of specific primers, two DNA fragments were amplified from FecB (190 bp) and exon 1 of GDF9 (462 bp) genes. The polymerase chain reaction (PCR) products were digested using AvaII and HhaI restriction enzymes for FecB and GDF9 genes, respectively. Results: In FecB locus, two genotypes, viz, wild type (+/+) and carrier-mutants (B/+), were detected with genotype frequencies of 0.948 and 0.052, respectively. Also, at GDF9 locus, two genotypes, namely, wild type (+/+) and carrier-mutants (+/-), were detected with genotype frequencies of 0.67 and 0.33, respectively. Heterozygous genotype frequencies for both loci showed higher than homozygous genotypes. None of the sheep carried homozygous genotype for both of the FecB and GDF9 variants in this breed. Conclusion: The results of the present study reveal slight polymorphism in FecB and GDF9 loci of Lori sheep breed. Keywords: Polymorphism, Lori sheep, Booroola gene (FecB), Growth differentiation factor 9 (GDF9), Polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP), Genotype, Mutants, Loci

Highlights

Improved animal genetic resources with versatile characteristics are necessary to confront growing human demand
Based on the methods described by Wilson et al [12] and Davis et al [17], forced polymerase chain reaction (PCR)–RFLP DNA test was used to detect the mutations of FecB and growth differentiation factor 9 (GDF9) in Lori breed sheep
Two genotypes, +/+ (190 bp), B/+ (190, 160 and 30 bp) for FecB and two genotypes +/+ (254, 156 and 52 bp), +/- (410, 254, 156, 52) for GDF9 were detected in Lori sheep (Figure 1 and 2)

Summary

Introduction

Improved animal genetic resources with versatile characteristics are necessary to confront growing human demand. Based on the methods described by Wilson et al [12] and Davis et al [17], forced PCR–RFLP DNA test was used to detect the mutations of FecB and GDF9 in Lori breed sheep. The result of chi-square analysis indicated that the population of Lori sheep for FecB locus was in Hardy-Weinberg equilibrium.

Results

Conclusion