The present study dealt with cloning and molecular characterization of MIC10 gene of T. gondii Kolkata isolate. TgMIC10 is a small microneme protein that lacks a putative transmembrane domain and appears not to remain associated with the parasite membrane after micronemal discharge. MIC10 gene of T. gondii “Kolkata isolate was PCR amplified and cloned into a TA cloning vector (Promega) to facilitate sequencing and subsequent characterization. The product revealed 597bp in size and has 99.2% homology with the published sequence. The difference was observed at 22, 210, 235, 378 and 513 positions. When the gene was deduced for amino acids, the homology was 98% with published sequence. The total amino acids number was 198 with predicted mass of 23 kDa molecular weight. The changes were observed at 8, 115 and 171th amino acid positions.