Polymorphisms in the kappa casein (CSN3) gene in horse and comparative analysis of its promoter and coding region

S. Hobor, T. Kunej, P. Dovc

Research output: Contribution to journalArticle

Abstract

The major parts of the coding region and promoter of the equine kappa casein (CSN3) gene were sequenced and compared among several species. Four SNPs were identified in the CSN3 gene: two in exon 1 and two in exon 4. The SNPs were genotyped in six Slovenian horse breeds using RFLP and two different PCR-based methods. The highest variation in genotype frequencies was found in the Slovenian cold-blood breed. The SNPs in exon 4 may cause a change in the amino acid sequence and may alter chemical/functional properties of the protein. Using horse-specific primers, we obtained 400 bp of exon 4 sequence from zebra and donkey. Two SNPs within the zebra exon 4 sequence were discovered; both presumably caused amino acid substitutions. Within the equine promoter sequence, 15 SNPs were found and 12 of them could be involved in the gain/loss of potential transcription factor (TF) binding sites. Using a comparative genomics approach, we obtained 1482 bp of the promoter sequence from zebra and donkey. Sequence alignment revealed highly conserved blocks of promoter sequence among nine species (sheep, goat, cow, zebra, donkey, horse, chimp, macaque and human) and clustered these species in three distinct groups. Consensus binding sites for TFs STAT5, C/EBP, NF1 and STAT6, previously demonstrated to be associated with expression, were located within conserved regions. Four promoter regions were tested for specific binding of TFs using electrophoretic mobility shift assays. Predicted binding sites for C/EBP and NF1 were confirmed and one conserved region was specifically detected by a yet-uncharacterized TF.

Original languageEnglish
Pages (from-to)520-530
Number of pages11
JournalAnimal Genetics
Volume39
Issue number5
DOIs
Publication statusPublished - Oct 2008

Fingerprint

kappa-casein
Equidae
Caseins
zebras
Genetic Promoter Regions
Horses
exons
promoter regions
genetic polymorphism
horses
Single Nucleotide Polymorphism
asses
Exons
binding sites
Genes
genes
Binding Sites
transcription factors
horse breeds
Transcription Factors

Keywords

  • CSN3
  • Electrophoretic mobility shift assay
  • Horse
  • Kappa casein
  • Promoter
  • Single nucleotide polymorphism
  • Transcription factor
  • Transcription factor binding site

ASJC Scopus subject areas

  • Genetics
  • Animal Science and Zoology

Cite this

Polymorphisms in the kappa casein (CSN3) gene in horse and comparative analysis of its promoter and coding region. / Hobor, S.; Kunej, T.; Dovc, P.

In: Animal Genetics, Vol. 39, No. 5, 10.2008, p. 520-530.

Research output: Contribution to journalArticle

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