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Kumar A.,Guru Jambheshwar University of Science and Technology | Chhokar V.,Guru Jambheshwar University of Science and Technology | Kumar R.,Guru Jambheshwar University of Science and Technology | Beniwal V.,Guru Jambheshwar University of Science and Technology | And 4 more authors.
International Journal of Pharma and Bio Sciences | Year: 2012

Mitochondrial Glycerol-3-phosphate acyltransferase (GPAM; EC 2.3.1.15), catalyzes the first step of glycerolipid biosynthesis. It plays a key role in the regulation of cellular triacylglycerol and phospholipid levels. The GPAM esterification step constitutes the committed reaction in the de novo synthesis of triacylglycerol (TAG) and all the acyl-glycerol phospholipids. In the present investigation GPAM was partially amplified (around 652 bp) by designing gene specific primers and confirmed by sequencing the amplicon and its comparison with the GPAM gene of bovine. Comparative study of GPAM among different breeds of buffaloes reveals different level of mutations with respect to its gene sequence 2.7-5.6% and protein sequence 3.4% to 13%. Similarly the protein structures modeled from their sequences were compared by structural superposition that shows variations (Root Mean Square Deviation) from 0.26 to 1.025. Molecular differentiation among different breeds of buffaloes reveals that the reference and Bhadawari are very close to each other, Toda and Surti are similarly close to each other and Pandharpuri is related to Murrah. Especially the variations are more at the binding site of this protein that may be the cause that different breeds have different percentage of milk fat. Further study is underway to detect polymorphism and associate them with milk fat related traits in buffalo.


Raut A.A.,High Security Animal Disease Laboratory | Kumar A.,Guru Jambheshwar University of Science and Technology | Kala S.N.,Central Institute for Research on Buffalo | Chhokar V.,Guru Jambheshwar University of Science and Technology | And 6 more authors.
Genetics and Molecular Biology | Year: 2012

Diacylglycerol O-acyltransferase 1 (DGAT1) is a microsomal enzyme that catalyzes the final step of triglyceride synthesis. The DGAT1 gene is a strong functional candidate for determining milk fat content in cattle. In this work, we used PCR-SSCP (polymerase chain reaction-single-strand conformation polymorphism) and DNA sequencing to examine polymorphism in the region spanning exon 7 to exon 9 of the DGAT1 gene in Murrah and Pandharpuri buffaloes. Three alleles (A, B and C) and four novel single-nucleotide polymorphisms were identified in the buffalo DGAT1 gene. The frequencies of the alleles differed between the two buffalo breeds, with allele C being present in Murrah but not in Pandharpuri buffalo. The allele variation detected in this work may influence DGAT1 expression and function. The results described here could be useful in examining the association between the DGAT1 gene and milk traits in buffalo. © 2012, Sociedade Brasileira de Genética. Printed in Brazil.

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