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Sporotrichosis is bipolar fungus Sporothrix schenckii induced skin, subcutaneous tissue and adjacent lymphatic subacute and chronic infections, mainly involving the limbs and face and other exposed parts, the performance of granulation tissue damage, occasionally involving mucous membranes, lungs, meninges, and other internal organs. Sporothrix is ??saprophytic fungi, widely distributed in the world, can contaminate soil, reeds, animal fur, sewage. Often due to trauma direct inoculation of human disease. Sporothrix schenckii as bipolar fungi, incubated at room temperature for the mycelial phase, the organization and incubated at 37 ℃ for the yeast phase. Its yeast phase can cause the skin, subcutaneous tissue, mucosa and local lymphatic system even systemic multi-system damage, and hyphal not. This phase pathogenic fungi and their closely related bipolar conversion. Currently, Sporothrix duplex conversion and pathogenic relationship between the two proteins has not been reported, while the other two-phase study fungi in this area there are many reports, many proteins have been speculated and identified. Studies have confirmed that there are differences in the yeast phase protein highly expressed, by comparison, our group will further study the Shen Keshi Sporothrix duplex conversion differential protein expression, and to look out for in the induction conversion mechanism plays an important role of the protein. Objective: To screen Sporothrix mycelial phase difference relative to the yeast protein, looking for information about bipolar conversion pathogenic proteins, as clearly Sporothrix pathogenesis and find targeted therapies lay an important foundation. Materials and Methods: standard strains of Sporothrix respectively hyphal and yeast phase of training, and their respective total protein extraction and purification. And identify suitable Sporothrix protein electrophoresis biphasic conditions for protein separation, protein profiles obtained related by Pdquest software analysis, select the relevant differentially expressed proteins after treatment for MS detection, and finally get relevant specific proteins . Results: 1, Shen Keshi Sporothrix after 25 ℃, can be seen in the light microscope slender mycelium into BHI medium 37 ℃ for 24 hours after cell breakage, 48 hours turned into a single spore . 2, during the two-dimensional electrophoresis Sporothrix, select the protein sample was 50μg, carrier ampholytes PH :4-6, by 2D clean-Up Kit fully purified parallel Coomassie blue staining for clear protein patterns for in proteomics research. 3, two-dimensional electrophoresis protein Sporothrix results measured over 300 different proteins, including the yeast phase of more than 200 high expression and low expression of more than 100. 4, by mass spectrometry and the NCBI database may be retrieved meaningful protein: DRK1 histidine kinase, heat shock protein 77 with 88, actin, cytochrome P-450, SU / serine kinase and some hypothetical proteins. Conclusions: 1 Sporothrix indeed appear in the temperature-dependent morphological transformation, 25 ℃ for the mycelial phase, 37 ℃ for the yeast phase. 2, the amount of protein in the sample and purification methods, the carrier ampholytes PH choice staining methods have affected the results of two-dimensional electrophoresis. 3, the establishment of Sporothrix mycelial phase yeast phase of two-dimensional electrophoresis patterns. 4, to obtain five possible pathogenic proteins: Hsp70 with Hsp88, SIR2 family of histone deacetylase, DRK1 histidine kinase, SU / serine protein kinase and phosphatidylinositol 3 - kinase.
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