The study aimed at characterization of buffalo β-casein gene and its promoter by PCR-SSCP analysis. Complete β-casein exon VII region analysis revealed two SSCP band patterns, with pattern-I representing predominant allele B (85%) present in homozygous (genotype BB) condition and pattern-II representing a rare allele A1 present in heterozygous condition (genotype A1B). Sequencing of two patterns revealed three nucleotide substitutions at codon 68, 151 and 193 of exon VII. The cDNA sequence of buffalo β-casein gene indicated three further nucleotide substitutions between allele A1 and B at codon 10, 39, and 41. Analysis of β-casein proximal promoter region (-350 upstream to +32) revealed four SSCP band patterns. These SSCP patterns corresponded to nucleotide substitutions at seven locations within 382 bp 5' UTR region of β-casein gene. Haplotype analysis suggested pattern-I of exon VII (wild type) was associated with three types of promoters and pattern-II of exon VII (rare type) corresponded to one exclusive type of promoter. The study suggested two haplotypes of exon VII and four haplotypes of promoter for buffalo β-casein.
Keywords: AP2; Beta casein; Buffalo; C/EBP; CCAAT/enhancer binding protein; E2F-myc activator/cell cycle regulator; EF2; Exon seven; MGF; Oct-1; PBS; PCR-SSCP; Phosphate buffer saline; Polymerase chain reaction-single strand conformation polymorphism; Polymorphism; Promoter; STAT5; UTR; Untranslated region; activating enhancing binding protein 2 alpha; mammary gland factor recognition sequence; nuclease factor octamer-1; signal transducer and activator of transcription 5.
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