Arsenic is a metalloid compound and it has become a severe threat to human health and environment when exceeding their maximum permissible limit of 0.01 mg/L in drinkingwater. Arsenic is a toxic and carcinogen substance which acts as an inhibitor for oxidativephosphorylation in our body and as a result, many diseases like skin cancer, stomach pain,kidney failure, cardiovascular disease etc can be found in human. So removal of arsenic from contaminated area is of great importance for human welfare. The main objective of this study was to functional analysis of arsC gene found in Lysinibacillus sphaericus which was capable of removing arsenic from arsenic contaminated LB medium. The Iterative Threading Assembly Refinement (I‐TASSER) tools were used to predict the secondary structure,molecular and biological annotation and 3D structure of arsC protein. Based on these analyses the function of this gene was predicted. The arsC gene was isolated from L.sphaericus and transferred to an arsC‐deficient strain of Escherichia coli MG 1655. RT‐PCR, colony PCR and blue white screening methods were used to confirm the insertion of this gene into this strain. Both transgenic and non‐transgenic strains of E.coli were exposed to arsenic stress and their tolerance and growth were studied. The transgenic strain was found to be more tolerant and could grow better when exposed to arsenics. These results suggest that the arsC gene of Lysinibacillus sphaericus B1‐CDA can be used as a potential candidate for genetic engineering to efficiently remove arsenics from contaminated sources like soil,water or industry effluents.