In earlier study a new thermophilic strain Geobacillus stearothermophilus strain-5 producing thermostable lipase was isolated and identified based on 16S rRNA sequencing. Phylogenetic analysis revealed its closeness to geobacilli especially the thermophilic Geobacillus stearothermophilus with optimal growth and lipolytic enzyme activity at 60°C and pH 7.0. In this study thermostable lipase gene from this bacterium was isolated by PCR using degenerate primers. The DNA fragment coding for lipase gene was cloned in the pCR 4-TOPO plasmid and the ligation products were transformed into Escherichia coli XL1-blue cells. Partial sequencing of the gene was carried out (accession number DQ923401). Analysis by BLAST program showed some sequence similarity to that, of several lipase genes from thermophilic Geobacillus and Bacillus submitted to Genbank.