Research Information System
1 st International Symposium on Graduate Research in Science Focus on Entrepreneurship and Innovation, İstanbul, Turkey, 4 - 06 October 2018, pp.60, (Summary Text)
Introduction
Pollens are the most important aeroallergens that cause allergy. The allergenic properties, chemical constituents and atmospheric concentrations of pollens change by climate, geographic location and environmental conditions. These factors lead to ambiguity in the diagnosis and treatment of pollen allergy. Today, recombinant allergens are being produced to cope with these problems. The main purpose of this study is to produce recombinant methionine synthase, a potential allergen found in local Morus alba (white mullberry) pollen, using fission yeast, Schizosaccharomyces pombe as host organism.
Materials and Methods
Total RNA was isolated from the pollens of M. alba, and cDNA library was constructed. The verification of the methionine synthase gene (metE) amplified by the polymerase chain reaction (PCR) from this cDNA library was conducted by the nucleotide sequencing. metE gene was ligated with a shuttle type vector, pSLF1073. Then this ligation product was transformed into S. pombe. After protein isolation from transformant colonies, recombinant protein was detected using Coomassie staining and Western blot methods.
Results
A protein band indicating metE protein of 85 kDa was observed in transformed cells both in Coomassie staining and Western blotting assays, while no band was detected in nontransformed cells. Thus, M. alba metE gene was successfully cloned in S. pombe for the first time in this study. It has been demonstrated that S. pombe can be used effectively in the production of recombinant allergens.
Innovation
Diagnostic kit and vaccine production studies can be carried out from the available data, and the experiences gained in this study will contribute to the development of national diagnosis kits for pollen allergies. Such efforts are expected to reduce cost and external dependency to the related pharmaceuticals, which are currently imported.
Keywords: Morus alba, methionine synthase, recombinant allergen.