lab of biochemistry division of influenza viruses, korea center for disease control and prevention...
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Lab of BiochemistryDivision of Influenza viruses, Korea Center for
Disease Control and Prevention
Mi-Seon Kim
Introduction
• What’s Candida albicans?
– Opportunistic fungal
pathogen of humans
– Invasion to internal organs
through the bloodstream
systemic infections
– C. albicans & Mortality rate
<Oral cavity infection>
Introduction• Previous work
– Target : Cell wall protein
• Why is the Cell-surface protein?
–More accessible to drugs than intercellular tar-
gets
• Cell surface protein– ALS1
– HWP1
CSF4
Procedure of Identification of cell-surface proteins in C. albicans
6 CSF 1~6
RT-PCR using primers inter-nal to the ORF
180 candidates
s
First allele replacement
Second allele replacement
First deletion cassette generated by 3-way PCR
Second deletion cassette
Target sequence
Target sequence
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)
Conclusion• Cell-surface proteins represent very attractive tar-
gets
• 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