Molecular characterization of Kathiawari horse using microsatellite markers
Raval, Ketankumar (2005) Molecular characterization of Kathiawari horse using microsatellite markers. Masters thesis, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand, Gujarat, INDIA.
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Genetic diversity can be described as genomic variation among individuals within and between breeds. Characterization of breed is the very first step for its conservation. Microsatellites have been found very useful as genetic markers for characterization. This study is an attempt to characterize Kathiawari breed of horse using six sets of microsatellite markers. A set of five bovine and six equine origin microsatellite primers viz.ILSTS- 005, ILSTS-011, ILSTS-030, ILSTS-033, ILSTS-034, LEX 025, LEX 027, LEX 032, LEX 034, LEX 035, and LEX 041 were tested for PCR amplification on randomly collected Kathiawari horse DNA samples. Out of these, six microsatellites viz. LEX 025, LEX 027, LEX 032, LEX 034, LEX 035, and LEX 041 were selected based on their ability to amplify chosen genomic regions. Out of fifty-three DNA samples, forty-six responded well to PCR amplification after optimisation of their quality. The PCR products were electrophoresed on 2% agarose along with 100bp DNA ladder to * S.D.A.V.. Sardarkrushinagar judge the size of different PCR products. The sizes of different PCR products were found to be approximately 145 bp, 195 bp, 260 bp, 250 bp, 252 bp and 155 bp for microsatellites LEX 025, LEX 027, LEX 032, LEX 034, LEX 035, and LEX 041 respectively. The amplified PCR products were further subjected to electrophoresis on 6% denaturing urea polyacrilamide gel (Denaturing PAGE) for allelic typing. Microsatellite LEX 025 exhibited five alleles in 6 different genotypic combinations. Allele # 3 was predominant in Kathiawari breed with maximum gene frequency (0.435). This allele was mainly expressed in heterozygous form. Allele # 1 was observed at least frequency (0.043). Microsatellite LEX 027 exhibited five alleles in nine different genotypic combinations. Allele # 2 occurred at maximum frequency i.e. 0.326. This allele mainly exhibited in heterozygous form. Allele # 1 occurred at minimum frequency Le. 0.044 which was observed in only 3 animals. Microsatellite LEX 032 exhibited five alleles in ten different genotypic combinations. Allele # 1 occurred at maximum frequency i.e. 0.348, which was mainly exhibited in heterozygous form. Allele # 5 occurred at minimum frequency Le. 0.054 which was observed in only 5 animals. Kathiawari horses exhibited four alleles in six different genotypic combinations at LEX 034 microsatellite locus. Allele # 4 occurred at maximum frequency (0.696), which was mainly exhibited in homozygous form. Allele # 3 occurred at minimum frequency i.e. 0.011, which was observed in only 1 animal. Microsatellite LEX 035 exhibited four alleles in eight different genotypic combinations. Allele # 1 occurred at maximum frequency (0.446). This allele mainly observed in heterozygous form. Allele# 4 occurred at minimum frequency (0.13). Kathiawari horses exhibited six alleles in eight different genotypic combinations at LEX 041 microsatellite locus. Allele # 5 occurred at maximum frequency of 0.402 in contrast to minimum frequency of 0.022 for Allele# 2 . The heterozygosity values at different loci were 0.684, 0.760, 0.755, 0.476, 0.702, and 0.717 for microsatellites LEX 025, LEX 027, LEX 032, LEX 034, LEX 035, and LEX 041 respectively. Microsatellite LEX 027 was highly polymorphic while microsatellite LEX 034 was the least polymorphic locus among all six microsatellites. The average heterozygosity of all six loci was 0.682. The polymorphic information content (PIC) was calculated from number of alleles and heterozygosity. The values were 0.621, 0.709, 0.704, 0.427, 0.645 and 0.662 for microsatellites LEX 025, LEX 027, LEX 032, LEX 034, LEX 035, and LEX 041 respectively. Microsatellite loci amplified in present study were found to be polymorphic with 4 to 6 alleles in Kathiawari horse. The heterozygosity and PIC ranged from 0.476 to 0.760 and from 0.427 to 0.709 respectively.
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