Cloning And Expression Of A New Allergen HSP7O Protein In Bombyx Mori And Establishment Of An Asthma Model

Sericulture in China has history of 5,000 years,extensive breeding of silkworm and the subsequent use of a large number of silk supplies make silkworm allergy reports often occur.Silkworms cause allergic clinical manifestations of allergic asthma,accompanied by rhinitis and conjunctivitis.In our study,we used proteomics and immunology for the first time to find silkworm HSP70 protein as silkworm allergen protein,and we successfully established mouse asthma model with recombinant HSP70 protein,which laid a good foundation for the specific treatment and diagnosis of allergy induced by silkworm.Purpose: using 2-DE and Western blotting method to found allergen in silk worm and provide evidence for the research of silkworm allergens and clinical diagnosis.Method:Total proteins of silkworm were separated by 2-DE,pooled sera from patients allergic to silkworm pupa proteins were used to detect IgE-binding proteins by western blotting;The positive proteins were further identified by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry(MALDI-TOF-MS)analysis;The cDNA of positive proteins gene was cloned and expressed in Escherichia coli;Allergenicity of the recombinant protein were investigated by ELISA and western blotting;An allergic asthma mouse model was also established with the recombinant protein as a specifc antigen.Results: eight potential allergens were identified by proteomics,and HSP70 is one of them.We susscessfully clone,express and purify the recombiant HSP70 protein.the recombinant protein also has a good allergenicity detected by ELISA and western blotting.Besides,Mice challenged with TP induced anobvious allergic asthma.Conclusion: HSP70 is a novel allergen from silkworm,it can induces allergic asthma,It provide evidence for specific prevention and treatment of allergic diseases.