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Title: PCR-SCCP and sequence analysis of leptin gene reveals novel polymorphism in intron 1 and allele fixation in exon 2 of Indian buffaloes (Bubalus bubalis)
Authors: Dubey, P K
Singh, Sandeep
Yadav, N K
Kathiravan, P
Mishra, B P
Kataria, R S
Keywords: Bubalus bubalis;Exon 2;Leptin gene;PCR-SSCP;Polymorphism
Issue Date: Jan-2014
Publisher: NISCAIR-CSIR, India
Abstract: The present study was undertaken to delineate single nucleotide polymorphisms within exon 2 and its flanking regions of bubaline leptin gene using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. A total of 118 buffaloes from seven different breeds were studied, revealing three distinct SSCP variants, viz., A, B and C, with a frequency of 0.703, 0.195 and 0.102, respectively. Sequence analysis of the region revealed 9 nucleotide variations including 8 transitions and one transversion as compared to cattle. Two A-G mutations within the coding DNA region were non-synonymous, resulting in change of amino acids. Basic local alignment search tool (BLAST) analysis of bubaline leptin gene revealed a sequence homology of 95-96% with bovines, caprines and ovines, while it varied from 86% to 93% with other farm animal species. Sequence analysis of SSCP variants showed a single nucleotide polymorphism (SNP) within intron 1 with the variant ‘A’ having homozygous ‘T’ allele and variant ‘B’ having homozygous ‘G’ allele, while variant ‘C’ having heterozygous ‘T/G’ alleles. The identified SNP locus was found to deviate from Hardy-Weinberg equilibrium with less than expected frequency of heterozygotes. The study also found no variation within exon 2 and complete fixation of unfavorable allele ‘C’, which has been earlier reported to be associated with lower milk and protein yield in cattle.
Page(s): 47-51
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.13(1) [January 2014]

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