Molecular characterization of HSP60 gene in Poonchi chicken
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Date
2022-01-17
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Sher-e-Kashmir University of Agricultural Sciences & Technology, Jammu (J&K)
Abstract
Heat shock protein 60 (HSP60) is a multifunctional protein, playing a crucial role in protecting organisms from environmental stress. The present study molecular characterization of HSP60 gene in Poonchi chicken was undertaken with the objectives of amplifying and sequencing of HSP60 gene and studying the genetic similarity and distance between different species to local chicken of Poonch. Total RNA was extracted from freshly collected chicken blood and cDNA was synthesized by reverse transcription. HSP60 gene was amplified from cDNA using specific primers designed by Primer 3 software. The amplified product was purified and thereafter cloned and sequenced. A partial cDNA sequence of chicken HSP60 gene of 1002bp encoding 327 amino acids was obtained. On its comparison, chicken partial cDNA sequence demonstrated 91.30%, 90.61%, 83.63%, 97.46%, 96.85%, 95.83%, 82.67%, 92.35% and 93.37% homology with pigeon, ostrich, rabbit, guinea fowl, turkey, quail, guinea pig, geese and duck, respectively. Deduced amino acid sequence of 327 residues of chicken HSP60 gene was 98.47%, 96.94%, 96.64%, 99.30%, 98.17%, 99.00%, 96.33%, 98.47% and 99.08% similar to pigeon, ostrich, rabbit, guinea fowl, turkey, quail, guinea pig, geese and duck, respectively. The phylogenetic tree showed conserved nature of HSP60 gene. Maximum divergence and minimum divergence were observed with guinea pig CDS and guinea fowl, respectively. In Z test it was found selection is of purifying type. It can be concluded from the present study that chicken HSP60, is highly conserved among different species and HSP60 gene might have been evolved by purifying selection (dS > dN).