Screening of Marwadi, Patanwadi and Dumba Sheep Breeds for Presence of Fecundity Gene Polymorphism by PCR – RFLP and Sequencing

Abstract

Mutation detection in major fecundity genes has been explored from the migratory flock of Marwadi,
Patanwadi and Dumba sheep of Gujarat. This study was conducted to assess whether any mutations in
BMP15 were exhibited in three different sheep breeds. The polymorphism of BMP15 was detected by RFLP
and SSCP. The FecXB locus amplified product of the BMP15 gene was digested using HinfI restriction
enzymes. The FecXL locus of the BMP15 gene was found to be polymorphic by SSCP followed by sequencing
of variant pattern. All the animals in the present study were found similar restriction fragment pattern,
among three genotypes-wild type (X++, 1.0) at FecXB locus. Hinf I site was not polymorphic in the present
sheep subpopulation. The nucleotide sequence of BMP15 gene, at FecXL locus in Marwadi, Patanwadi &
Dumba sheep was also searched for polymorphisms by PCR-SSCP and investigated for variants using
cloning-sequencing. The nucleotide sequences were subjected to alignment analysis and it was observed
that among three, two nucleotides change in the bases were found, one at position 53, C-T base change and
the second at position 182, C-T base change compared to the reference sequences. Out of total 60 sheep
screened for mutation for BMP15 gene, all animals were found homozygous wild type for BMP15 gene for
PCR-RFLP. None of the animals were found homozygous mutant or heterozygous (carrier) type for BMP15
gene. SSCP-sequencing results showed that the two nucleotide changes might be related with the fecundity
of sheep. The present findings indicate that the three sheep populations are not carriers for BMP15 gene
mutation.