MADRAS AGRICULTURAL JOURNAL

Molecular cloning and characterization of venom peptides from two Honeybee species (Apis cerana and Apis mellifera)

Abstract

                                ABSTRACT Two honey bee species (Apis cerana and Apis mellifera) are domesticated for the economic products likehoney,royal jellyandvenom. Three components ofhoney bee venom peptide viz., melittin, phospholipase A2 (PLA2), and secapin have proven effects in pharmaceutical and clinical applications and these components were cloned and characterized in the present study. Bee venom glands were dissected out for RNA isolation and cDNA was synthesized from the total RNA. PCR was carried out using gene-specific primers for melittin, PLA2,andsecapingenes,using cDNAas template.PCRproducts were cloned in the TA-cloning vector and sequenced. Amino acid sequences were deduced from nucleic acid sequences and a phylogenetic tree was constructed to deduce the evolutionary relationship among the honey bee species.Crude bee venom was collected by electric milking method and dissolved in sterile water to separate the venom peptides by SDS-PAGE. Crude bee venom was tested against the rice pathogenic bacterium Xanthomonas oryzae pv. oryzae which causes bacterial blight disease of rice. Agar diffusion assay of bee venom was performed to confirm the antimicrobial properties of crude venom. Both A. cerana and A. mellifera venom showed antibacterial activity against X. oryzae pv. oryzae.

Key words : RT-PCR, Bee venom peptides, Melittin, Phospholipase A2, Secapin, antibacterial activity