Sequencing of 120 Mycobacterium tuberculosis clinical isolates for gyrA gene revealed that 109 are having change of amino acids that resulted in no physiological change or the structural change in the protein. However, bioinformatics analysis of docking with the ligand Ofloxacin revealed that there are minor changes in binding energies of changed sequences that may have some physiological impact in terms of its expression or may have evolutionary significance to be studied in future work.