HSP90gene of O.niloticus sp. is an associate of the heat shock protein family and plays a significant role in stress-related and defense responses educed by contagion with activator of hsp90 ATPase activity. Hence, understanding molecular structure and function of the protein coded by this gene is of paramount importance for fish biologists working on ATPase. The present study was aimed at sequence and in silico structural analysis of Hsp90 protein coded by this gene, through Validation of the overall folds and structure analysis 36%, Alpha helix,17% beta strand,38% in normal mode. One major domin were detected belonging to hsp90 family while neural network analysis is related protein to be highly phosphorylated at threonine residues. Sterochemical analysis shows 80.5% of most favoured region.16.4% additional allowed region 2.4% in generously allowed regions.