In the present study, out of four alleles amplified from seabass (Lates calcarifer) genome inhabiting Mumbai water by PCR using growth hormone (GH) gene-specific primers, two DNA fragments (SGMS1, 233 bp and SGMS2, 239 bp) were eluted from gel, cloned using pTZ57R (2.886 kb) vector into E. coli DH5α, characterized by restriction endonuclease analysis and sequenced by automated DNA sequencer. After blasting and multiple alignment of the above sequences, SGMS1 showed 97% and SGMS2 93.3% homology with promoter region of GH gene containing microsatellite of Australian seabass and 94.6% homology between both the fragments. These sequences SGMS1 and SGMS2 were submitted to NCBI GenBank. On blasting, these sequences with gene databases, SGMS1 and SGMS2 showed partial homologies with Seriola quinqueradiata (26.9%, 12.9%), flounder (15.8%, 15.8%), Oreochromis nilotica (23%, 7.9%), Oreochromis mossambicus (23%, 7.9%) and Danio rerio (8.2%, 7.5%). Critical analysis showed the presence of microsatellite (CA)16 and (CA)19 repeats in fragments SGMS1 and SGMS2, respectively in seabass from Mumbai water in comparison to (CA)14 repeats from the Australian seabass. Further, on sequence comparison, single nucleotide mismatches detected at their several positions in relation to seabass GH gene of Australia. These nucleotide variations detected in SGMS1 and SGMS2 in comparison to those of the Australian seabass may be due to mutations owing to environmental or habitat changes that seem to have definite potentials for development of genetic markers, which would be useful for identification and selection of superior germplasm with desirable commercial traits such as high growth rate.