To predict the three dimensional structure of testis anion transporter-1(TAT1) protein by using I-TASSER tool. TAT1 protein sequence was collected from different tools like SWISSPROT, PDB. Characterization was performed for TAT1 protein with Ramachandran plot, PyMOL, Rasmol. Sample size was calculated using clinical.com with previous literature (N=10) times repeated for C score, TM score and RMSD. Results: 3D model structure of TAT1 protein was visualized by PyMOL. Psi and phi angles of testis anion transporter-1 protein was performed by Ramachandran plot. C-score, estimated TM-Score and estimated RMSD of protein were -1.15, 0.57±0.15, 11.6±4.5 respectively. Significance was observed for the comparison of parameters like C-score, estimated TM score, estimated RMSD through SPSS Version 2.1 One sample T test with P(0.001) reveals statistical significance. The three dimensional structure of testis anion transporter-1 protein was predicted by I-TASSER. It could be used as a biomarker for male infertility diagnosis. Further the characterization of TAT1 protein leads to development of crystalline structure. [ABSTRACT FROM AUTHOR]