Allergy is one of major health problems of today. One of the most important aero allergens which causes allergic sensitivity and allergic disease symptoms is pollens. The allergenic properties, chemical constituents and atmospheric concentrations of pollens change by climate, geographic location and environmental conditions. These factors lead to some problems in the diagnosis and treatment of pollen allergy. Nowadays in order to overcome these problems, recombinant DNA technology is used to produce recombinant allergens that mimicnatural, functional, and immunologically natural allergens, at a constant quality and sufficient quantity. The main purpose of this study is recombinant production of methionine synthase, a potential allergen found in local Morus alba (white mullberry) pollen, in Schizosaccharomyces pombe. Firstly, total RNA isolation was performed from the pollen of M. alba, and a cDNA library was set up. The methionine synthase (metE) gene was amplified by the polymerase chain reaction (PCR) from this cDNA library and the complete gene sequence was determined by nucleotide sequencing. metE was ligated with a shuttlety pevector, pSLF1007 and this ligation product was transformed to S. pombe. Protein isolation was performed from transformant colonies and Western blot analysis was performed to detect the recombinant protein. The recombinant protein was detected on the membrane by using an antibody specific to the histidine tag. As a result, metE found in M. alba pollen was successfully produced in S. pombe recombinantly. In this study, S. pombe has been shown to be an alternative system for the production of recombinant allergens. Recombinant methionine synthase has a potential to be used further clinical studies, and to become a biotechnological product in the market after purification and approval of it saller genic properties.

Keywords: Gene cloning, Methionine synthase, Morus alba, Recombinant allergen, Schizosaccharomyces pombe