Molecule cloning of a major allergen acid phosphatase (Apim3) gene from honeybees and construction of a prokaryotic expression vector.

Objective To clone a major allergen acid phosphatase(Apim3) gene from honeybees(Apis cerana cerana) and construct a prokaryotic expression vector.Methods Special primers were designed on the basis of the reported gene.Total RNA was extracted from bees.RT-PCR was used to amplify the Apim3 gene and sequence analysis was performed.Fragments were subcloned into the expression vector pET-28a.Results The A pim3 gene was obtained from honeybees and a prokaryotic expression vector was constructed.Analysis revealed that the gene consisted of 1 167 nucleotides encoding 388 amino acids with a molecular weight of 45 279.9 and pI of 5.53.The gene had homology of 97% with the gene reported in GenBank.Conclusion The honeybee Apim3 gene was cloned and a prokaryotic expression vector was successfully constructed.This should prove to be of considerable significance in determining the expression of allergens and immunological activity.