Molecular characterization of Zalawadi goat using microsatellite marker
Thakkar, Kamlesh (2002) Molecular characterization of Zalawadi goat using microsatellite marker. Masters thesis, College of Veterinary Science and Animal Husbandry, Anand Agricultural Univeristy, Anand, Gujarat, INDIA.
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Genetic variation between and within breed is described as diversity. It is essential to characterize a breed for its conservation. Microsatellites are ideal molecular markers for characterization. Present study is an attempt to characterize Zalawadi goat bred using fice sets of bovine origin microsatellite markers. A set of ten bovine origin microsatellite primers viz. ILSTS 005, ILSTS 006, ILSTS 011, ILSTS 030, ILSTS 033, ILSTS 034, ILSTS 037, ILSTS 044, ILSTS 049 and ILSTS 072 were tested for PCR amplification from goat DNA samples. Out of these ten microsatellites, five microsatellite primers viz. ILSTS 005, ILSTS 011, ILSTS 030, ILSTS 033 and ILSTS 034 were chosen based on their ability to amplify goat microsatellites. Fifty DNA samples were subjected to PCR amplification of these microsatellites. All samples did not respond to PCR amplification. Fourty three to 49 samples yielded amplification. The PCR products were checked on 2-2.5% agarose for amplification and the positive samples were run on 2% metaphore along with 100 bp DNA ladder to judge the size. The sizes (lengths) of different microsatellites were found to be approximately 183 bp, 260 bp, 148 bp, 151 bp and 140 bp for microsatellite ILSTS 005, ILSTS 011, ILSTS 030, ILSTS 033 and ILSTS 034 respectively. Further, these amplified products were run on 4% agarose for 2-3 hrs for preliminary screening. Further the amplicons were run on sequencing PAGE (Denaturing PAGE) for allele typing. Microsatellite ILSTS 005 showed four alleles in eight different genotyping combinations in Zalawadi goat. Allele 3 had highest frequency (0.445) while allele 2 has the least 0.022 in this breed. Microsatellite ILSTS 011 exhibited four alleles in nine different genotypes. Allele #2 occurred at highest frequency (0.667) mainly exhibited in homozygous form. While allele #3 at lowest (0.104). Microsatellite ILSTS 030 was having 7 alleles in 17 different genotypic combinations. Allele 2 and 4 constituted more than of total 0.60 frequency. Microsatellite ILSTS 033 showed 7 alleles in ten different genotype. Although there are seven alleles, only two alleles, #4 and #5 constituted more than 0.75 of total frequency. Microsatellite ILSTS 034 expressed only three alleles in four different genotypic combinations. Allele #2 and #3 constituted more than 95% of total frequency. The heterozygosity values in these microsatellites were 0.554, 0.524, 0.753, 0.692 and 0.512 for microsatellite ILSTS 005, ILSTS 011, ILSTS 030, ILSTS 033 and ILSTS 034 respectively. Microsatellite ILSTS 030 was highly polymorphic and ILSTS 034 was least polymorphic among all five microsatellites. Polymorphic information content (PIC) was calculated from number of alleles and heterozygosity. It was 0.496, 0.484, 0.705, 0.639 and 0.398 for microsatellite ILSTS 005, ILSTS 011, ILSTS 030, ILSTS 033 and ILSTS 034 respectively. Five goat microsatellite were amplified using bovine microsatellite primers. The microsatellites were found to be highly polymorphic with 3 to 7 alleles, 0.512 to 0.753 heterozygosity and 0.398 to 0.705 PIC in the Zalawadi goat. This is the first reported molecular characterization study in Zalawadi goat.
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