Genotyping Betacasein Gene (Exon 7) in Sahiwal Cattle by PCR-SSCP
Journal Title: Journal of Animal Science Advances - Year 2012, Vol 2, Issue 1
Abstract
Casein genetic polymorphisms are important and well known due to their effects on quantitative traits and technological properties of milk. At the DNA level, polymerase chain reaction–single strand conformation polymorphism (PCR-SSCP) allows for simultaneous typing of several alleles and in the detection of unknown polymorphisms. Sahiwal is an indigenous dairy breed of cattle, yet little is known about the DNA polymorphism of the milk protein genes. Sahiwal cattle (n=196) were typed by PCR-SSCP. The amplified products were denatured and run on 12% non-denaturing gels followed by silver staining. Nine SSCP gel patterns were observed which were based on the number of bands and in the shift in their mobility. Sequencing of the 234 bp fragment of β-casein (exon 7) followed by edition to 137 bp reduced the nine different patterns to three variants based on the presence of two single nucleotide polymorphisms (SNP’s). Comparison of variant I (54.08 %) with the reference sequence (Gene Bank Acc. No. M55158) identified a synonymous SNP (C→T) at position + 8261 with a deduced aminoacid of threonine. Variant II (44.90%) was similar to the reference sequence. Variant III (1.02 %) revealed a transversion from CG at position +8267 of the reference sequence with a deduced aminoacid change of Serine (C)Arginine (G) and corresponding to codon 122 of the reference protein β– CN A2-5P. This may partly be attributed to the presence of the B allele in the population studied.
Authors and Affiliations
L. Dhanammal, I. D. Gupta, S. Rachagani, A. Verma
Keywords
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