Vol. 12, Special Issue 8 (2023)

Author(s):
Harpreet Singh, Komal deep Kour, Nishant Kumar, Mandeep Singh Azad and Palak Sharma

Abstract:
Heat shock protein 70 (HSP70) perform many vital functions in the cells and is responsible for cytoprotection under stressfull conditions. In this study, molecular characterization of HSP70 gene in local chicken of Poonch was undertaken with the objectives of amplifying and sequencing of HSP70 gene and studying the genetic similarity and distance between different species with respect to local chicken of Poonch. Fresh blood was collected from chicken and used for RNA extraction. cDNA was synthesized by reverse transcription. HSP70 gene was amplified using specific primers designed by Primer 3 software. The amplified product (1986 bp) was purified and ligated in pGEM T Easy vector. The ligated product was cloned using DH5α competent cells and then sequenced. MEGA X and DNA STAR softwares were used for sequence analysis of the HSP70 gene. The comparison of HSP70 CDS sequence demonstrated 100%, 95.96%, 95.85%, 95.70%, 92.60%, 91.44% 91.39%, 91.39%, 81.47%, 81.41% and 81.36% similarity with chicken, quail, turkey, guinea fowl, pigeon, ostrich, emu, duck, goat, sheep and pig, respectively indicating sequence homology among different species was high. The percentage identity of translated protein sequence of 634 residues of HSP70 gene was 100%, 98.26%, 98.11%, 98.11%, 97.95%, 97.95%, 97.48%, 97.32%,92.36%, 91.82% and 91.51% similar to chicken, quail, turkey, guinea fowl, pigeon, duck, emu, ostrich, pig, sheep and goat, respectively. The phylogenetic tree drawn by MEGA X software showed that the gene is evolutionarily conserved. Z test was conducted which showed that the gene might have evolved by purifying selection (dS>dN).