Research Of A Kind Of Allergenic Storage Protein From Tartary Buckwheat

The buckwheat has higher nutritional quality and medical value, and includes many kinds of bioactive material richly. These substances have shown significant efficient in preventing and curing the modern civilization diseases, cardiovascular diseases and rheumarthritis such as diabetes, hypertension etc. But the anaphylaxis compositions contained in the buckwheat cause a lot of person who contact or eat it produce allergy symptom. In the last few years, some foreign scientists have studied the allergic composition in common buckwheat (CB). But reports in allergic composition research of tartary buckwheat (TB) are rare, it still belongs to the blank that the research about tartary buckwheat allergenic storage protein.According to the amino acid sequence of TBa purified from tartary buckwheat seeds which was sequenced by MALDI-TOF-MS, and nucleic acid sequences of allergenic protein from common buckwheat, the primers were designed. Using method of PCR, 3' and 5'- RACE, the full length cDNA sequence coding of an allergenic protein in tartay buckwheat (TBc) was amplified. The sequence consists of 1773 bp nucleic acids. 5' end sequence of 45bp contain GC richly, there is a open reading frame among 46 to 1593bp, which encoding 515 amino acids, 3' end include 200bp. TBc was made up of two subunits, TBa and TBb. TBa includes 195 amino acid residues, TBb includes 320 amino acid residues. Data analysis of nucleotide acids and deduced amino acids is performed, including conservation analysis of its amino acids, prediction of position of disulfide bridge and the location of post-translation, signal peptide prediction, second structure and 3D structure prediction, phylogenetic analysis, antigen sites prediction.TBa, TBb and TBc are purified respectively, with salting out, anion exchange chromatography, gel filtration and rapid efficient electrophoresis elution, and then immunological character of three proteins are analyzed.According to the above experimental result, we infer the tartary buckwheat allergenic storage protein (TBc) is composed by two subunits, Tba (MW...