Preparation of recombinant most abundant protein MRJP1 of royal jelly

The cDNA encoding major royal jelly protein MRJP1, which is the most abunda nt protein of royal jelly (RJ), was cloned to pQE32 vector without most of the signal peptide sequence. The recombinant His-tag protein (rMRJP1) after expression was purified under denaturing conditions following purification by affinity chromatography on Ni-NTA resin. Since the rMRJP1 after affinit y purification was contaminated with a 30 kD protein of E. coli which was n ot possible to remove by any QIA express wash and elution procedure, pure r MRJP1 was obtained by electroelution from the polyacrylamide gel. The rMRJP 1 will be used for testing of allergic reactions of patients allergic to ro yal jelly and honey to confirm its allergenicity described recently for the main 55-57 kD RJ and honey protein as well as to detect the role of both t he amino acid sequence or the glycosylated moieties of MRJP1 in the inducti on of allergy.