DNA gyrases, classified as type II topoisomerases, are critical for regulating bacterial DNA topology and serve as primary targets for fluoroquinolone antibiotics. Resistance to these antibiotics frequently arises from mutations in the gyrase subunits, particularly GyrA and GyrB. This study addresses the structural characterization of the GyrB subunit of Helicobacter pylori DNA gyrase, whose tertiary structure remains experimentally uncharacterized. Utilizing homology-based modeling via the Swiss-Model server, we predicted the 3D structure of GyrB, leveraging sequence data from public databases (NCBI, Pfam, UniProt). Conserved functional domains—HATPase_c (ATPase catalytic domain), DNA_gyrase_B, and Toprim—were identified, with sequence alignments revealing high evolutionary conservation across bacterial species. Structural annotation further localized critical residues within these domains, including catalytic sites (e.g., ATP-binding motifs) and regions associated with fluoroquinolone resistance. Notably, mutations proximal to the Toprim domain, implicated in stabilizing the DNA-enzyme complex, were mapped to residues previously linked to quinolone resistance in other pathogens. These predictions provide the first computational structural framework for H. pylori GyrB, elucidating potential mechanisms underlying drug resistance. The model underscores the utility of homology-based approaches in bridging structural knowledge gaps for antimicrobial targets. Our findings establish a foundation for targeted mutagenesis studies and structure-guided drug design to combat fluoroquinolone-resistant H. pylori strains, emphasizing the need for experimental validation to corroborate these insights.