Homeobox genes encode transcription factors which play important roles in the developmental processes of many multicellular organisms. TGIFLX/Y (TGIFLX and TGIFLY) are members of the homeobox superfamily of genes. Their expressions are specifically detected in the human adult testis but their functions are remained to be investigated. In this investigation we cloned full length of TGIFLY cDNA and produced recombinant GST-TGIFLY protein in bacterial system. Here we present production of GST-TGIFLY fusion protein as a soluble protein. The recombinant protein was confirmed by western blot analysis using anti-GST antibody. Through a single purification procedure using MagneGST Beads, approximately 20 mg of the recombinant protein was obtained per liter of bacterial culture. We suggest that GST-TGIFLY fusion protein could be utilized as a valuable molecular tool on investigation of TGIFLY target genes and identification of co-factors or partner proteins involved in TGIFLY function in normal and abnormal development.