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Lab of Biochemistry Division of Influenza viruses, Korea Center for Disease Control and Prevention

Lab of Biochemistry Division of Influenza viruses, Korea Center for Disease Control and Prevention Mi- Seon Kim. Contents. Introduction Identification method In vitro experiments In vivo experiments Conclusion. Introduction. <Oral cavity infection>. What’s Candida albicans?

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Lab of Biochemistry Division of Influenza viruses, Korea Center for Disease Control and Prevention

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  1. Lab of Biochemistry Division of Influenza viruses, Korea Center for Disease Control and Prevention Mi-Seon Kim

  2. Contents • Introduction • Identification method • In vitro experiments • In vivo experiments • Conclusion

  3. Introduction <Oral cavity infection> • What’s Candida albicans? • Opportunistic fungal pathogen of humans • Invasion to internal organs through the bloodstream  systemic infections • C. albicans & Mortality rate

  4. Introduction • Previous work • Target : Cell wall protein • Why is the Cell-surface protein? • More accessible to drugs than intercellular targets • Cell surface protein • ALS1 • HWP1

  5. Domain organization of a typical fungal cell-wall protein Glycosylation

  6. Procedure of Identification of cell-surface proteins in C. albicans 180 candidates 6 CSF 1~6 RT-PCR using primers internal to the ORF CSF4

  7. Primary sequence analysis of C.albicans Csf4 protein < CSF4 - gene product of ORF6.1639 >

  8. First deletion cassette generated by 3-way PCR Target sequence s First allele replacement Target sequence Second deletion cassette Second allele replacement

  9. Filamentation assay Clinical isolation Parental strain Mutant • Invasion upon Agar • Filamentation(Hyphae formation) • Growth on the Agar surface • Severe defect of Filamentation • No filaments around colony • Targeted gene =No growth factor • Viable csf4 null mutant • Similar growth rate (csf4 null mutant & wild type)

  10. Adherence assay Wild- type • Support a role for Csf4 in the adhesion of C.albicans to mammalian cell

  11. Virulence Assay in vivo Uninfected Csf4 null_HC SC5314 BWP17_ARG/HC Csf4 null_HC SC5314 als1 null_HC

  12. Virulence Assay in vivo • CSF4 • Factor of Rapid invasion • Destrction of key organ

  13. Conclusion Cell-surface proteins represent very attractive targets Csf4 belongs to the familly of yeast glycosidases Csf4 is important for adhesion of C.albicans to mammalian cells in vitro Csf4 is good candidates for new virulence factor Csf4 is represents a very promising target for the development of antifungal drugs

  14. Thank you

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