PCR - SSCP and nucleotide sequencing of k-casein gene in buffalo.

摘要

The study was conducted to determine the genetic polymorphism in exon 2 region of kappa -casein gene using PCR-SSCP. The 112 bp gene fragment of kappa -casein composed of partial intron 1, exon 2 and partial intron 2 were amplified by PCR. The nucleotide sequencing of different alleles identified by PCR-SSCP was performed using automated DNA sequencer. Results revealed that two SSCP patterns (AA and AB) were observed in the four breeds of buffaloes. Genotypes AA and AB with genotypic frequencies of 0.9 and 0.10 were observed in Murrah buffalo, respectively. Bhadawari had AA and AB genotypes with genotypic frequencies of 0.92 and 0.08, respectively. Genotype AA was observed in both Surti and Mehasana. Two new alleles A and B were identified. It was observed that A allele had AGG at thirty-ninth position of 112 bp fragment, whereas AGT was present at this site in cattle and buffaloes. B allele had GGT at the fiftieth position, whereas GTT was present at this site in the case of cattle and buffaloes.