Bee venom is a colorless acidic liquid and can cause an anaphylactic reaction in higher doses. However, the therapeutic potential is enormous in a lower concentration, including anti-cancer, anti-microbial, anti-inflammatory, and anti-oxidant properties. Bee venom contains very few pharmacologically vital substances, and melittin is the principal component besides phospholipase A2, histamine, hyaluronidase, catecholamine, and serotonin. Melittin has demonstrated a range of therapeutic benefits in preclinical cell culture and animal models, including anticancer, anti-bacterial, anti-protozoan, and immunomodulatory properties. In the present study, we investigated physicochemical composition, protein secondary structure, and antigenicity of the melittin peptide from Apis cerana indica through in silico approach and investigated the influence of melittin intron for transgene expression in the mammalian expression host system. From our study, we found that the mature melittin peptide from A. cerana indica is highly unstable, basic, and cationic in nature and the melittin intron has no positive effect on transient expression of exogenous gene in HEK-293 and CHO-K1 cell line.