Thermal pressure alters the normal body homeostasis and cause rigorous injurious sound effects on production of cattles. Dairy cattles are extra responsive to high temperature as they spawn further energy expanded rate and this matter grasp particular consideration when the entire picture of inclusive warm up is measured around the globe. Heat fright proteins are ever-present, highly conserved proteins that cooperate a significant task in cell endurance through unusual stresses. The present study was undertaken to assess peculiar relevance of HSP70 gene with heat tolerance ability in Red Sindhi (Bos indicus), tropically evolved breed of Pakistan. The aspiration of this study was to explore and portray the heat shock proteins gene diversity in coding region of Red Sindhi cattle to understand their thermotolerance under hot conditions using Polymerase Chain Reaction (PCR). The complete coding region of HSP70 is 1926bp long without any intron, programming a protein of 641 amino acids located on chromosome No.23, with predicted molecular weight (Mw) of 70,190.56 kDa. The functional parts of HSP70 protein consist of an amino-terminal or N-terminal 44kDa ATPase domain (amino acid residues 1-386), 18kDa domain is peptide binding (a.a 384-543) and highly conserved 10kDa Carboxyl or C-terminal domain (residues 542-640) in all eukaryotes. The phylogenetic examination of mammalian HSP70 gene indicated that Red sindhi has showed (99.79 to 99.48%) homology with Bos taurus, Bubalus bubalus and Capra hircus while least similar relationship was observed with Canis lupus that was 86.34%. The results express that nucleotide as well as deduced amino acid order was greatly preserved. Comparative sequence analysis of Red sindhi HSP70 gene revealed a total of 14 single nucleotide polymorphic sites, of these 5 were non-synonymous (C14T, G970T, C1469T, A1535T, G1560T) caused change in amino acid sequence while 9 were synonymous. In non-synonymous SNPs the 1 one was reported and four novel type. These 9 mutations caused no change in amino acid. 8 out of 14 were transition type in which purines were replaced by just purines and pyrimidines replaced by other pyrimidines. 6 types were transversion mutations in which purines replaced by pyrimidines and vice versa. At protein level, Red sindhi HSP70 gene has showed total 5 amino acid changes and two of five are in N-terminal domain (M5T, A324S), remaining are in peptide binding domain (A490V,K512M, Q520H). Probably damaging functional impact of amino acid change is seen in two of expressed SNPs. These transformations can be worn as molecular genetics pointer to aid assortment for anti-heat stress cattles. The findings of this study revealed that detected polymorphisms may be associated with ability of bovine to adapt with tropical and harsh climatic conditions but additional exploitation still required to draw the ending about molecular roots of thermotolerant cattles.