Sequence characterization and SNP identification of TNP1 gene in Indian cattle breeds

Abstract

Present study was conducted on 50 bulls and 40 male calves of Sahiwal, Tharparkar and Karan Fries cattle maintained at ABRC and LRC, NDRI Karnal (Haryana) to characterize and identify genetic polymorphisms in TNP1 gene. A total of 1568 bp region of TNP-1 gene includes 490 bp of promoter region and two exon and one intron was sequenced and characterized in Bos indicus cattle breeds which are widely distributed in Indian sub-continent. Four sets of primers for TNP1 gene on the basis of Bos Taurus sequence (Acc. No- BK_006511) were designed using Primer3 software and PCR products of 487, 450, 455 and 250 bp were obtained. Amplicons were custom sequenced and subjected to Clustal W analysis which showed no nucleotide changes in coding region and non coding region in Indian cattle breeds as compared to Bos taurus. The 490 bp of promoter region was subjected to transcription factor binding site. Three TATA boxes and two CAAT boxes were identified in the studied fragment. Analysis of SNP was performed using restriction fragment length polymorphism (PCR-RFLP), to detect nucleotide changes in the sequence as reported (g.528G>A, SS1388116558) in Chinese Holstein breed. No polymorphisms were found for tested SNP. Only one genotype GG indicates the absence of variability in the sampled population.