GENETIC CHARACTERIZATION OF NATIVE CHICKEN OF HIMACHAL PRADESH USING MOLECULAR MARKERS

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Date
06-07-11
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CSKHPKV, Palampur
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ABSTRACT Sixty five (65) blood samples at random from birds procured from different locations of the state and maintained at CSKHPKV Poultry farm, Palampur were collected. Fifteen microsatellite markers (MCW073, MCW005, MCW004, MCW051, MCW049, MCW048, MCW001, MCW014, ADL-158, ADL-172, ADL-267, ADL-210, ADL-171, ADL-176, ADL136) which gave amplification were included in the analysis. The PCR products for different microsatellite loci were resolved on 6% denaturing (urea) polyacrylamide gels along with 50 and 100 bp DNA ladders at 40-45V. Microsatellite alleles were visualized by silver staining. The observed alleles, effective number of alleles, gene frequency, observed heterozygosity, expected heterozygosity were estimated through POPGENE software. The mean observed and effective numbers of alleles were found to be 5.87 ± 0.28. and 4.19 ± 0.17 respectively across all loci. Total 88 alleles were observed with maximum alleles (8) contributed by locus MCW073, ADL-176 & and the least alleles (4) by (ADL-171, ADL-136, MCW014). The average heterozygosity was observed to be 0.76 ± 0.01 across all loci. The average observed and expected heterozygosity 0.56 ± .03 and 0.77 ± 0.01 respectively. The observed heterozygosity (0.8900) was highest for ADL-176, and least (0.4082) for ADL-267. The Expected heterozygosity (0.8278) was highest for MCW073 and least (0.6890) least for MCW014. All the microsatellites had shown high levels of genetic variability.The screened loci were not found to be in accordance with Hardy-Weinberg equilibrium for several microsatellite frequencies. Genes ChB6 and IL-2Rγ were used to study nucleotide polymorphism related to immunocompetence The PVU-II restriction enzyme digestion of 215bp amplicon of ChB6 gene generated fragments of 215,147 & 68 bp and revealed highly polymorphic patterns. The Hph1 restriction enzyme digestion of 600bp amplicon of IL-2Rγ gene generated fragments of 465,454,134,104,42bp and revealed highly polymorphic patterns. Fragment pattern revealed all the genes have high level of polymorphism in DNA samples of Native Chicken of Himachal Pradesh.
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