Molecular Cloning and Characterization of a Venom Phospholipase A2 from Apis mellifera spp.

Abstract

Honeybee venom plays a central role in the biodiversity and self-defense of honey bee, which protect them by allowing the stinger to immobilize, puncture, and initiate allergy on their predators. Phospholipase A2 (PLA2) is one of the main allergic enzymes of honeybee venom, and it has been extensively studied in biochemical properties due to the toxicity to insects and other invaders causing hemolysis on erythrocytes. The present study described the results of cDNA cloning and chemical characterizations analysis of venom PLA2 from honeybee, Apis mellifera spp., an economically important insect for pollination on crops. The gene encoding Phospholipase A2 from honeybee (A. mellifera spp.) venom was isolated, cloned and its nucleotide sequence was determined. The nucleotide of PLA2 is 507 bp in length, and shared 99% and 96% homology with those of Apis mellifera ligustica and Apis cerana cerana, respectively. The amino acid sequence deduced of PLA2 resembled those of other bee subspecies, such as western honey bee (A. mellifera lingustica). Predicted amino acid sequences of PLA2 is 99% identical with that of A. mellifera lingustica, and its pI is 7.01 and lower than those of A. mellifera lingustica and A. cerana cerana. In order to understand better the phylogeny of PLA2, we analyzed its nucleic acid sequence and compared with those of other sources from NCBI database, and modeled its 3D structure based on template d1poca. Amino acid sequences of PLA2 from related species were obtained to construct a phylogenetic tree and revealed that PLA2 of A.mellifera spp. was belong to group of the western honey bee, and was most closely related to that of A. mellifera ligustica than other bee subspecies.