CLONING OF A DNA FRAGMENT ENCODING PART OF A 70-KDA HEAT-SHOCK PROTEIN OF CANDIDA-ALBICANS

Immunoscreening of a mycelial expression library with polyclonal antib odies raised against mycelial cell wall resulted in the detection of a cDNA encoding a heat shock protein of Candida albicans. Sequence anal ysis of a 0.8-kb cDNA subclone, 2M-1, revealed an open reading frame e ncoding 244 amino acids. Southern blot analysis with this fragment as a probe demonstrated hybridization to C. albicans DNA. Northern analys is showed a substantial increase in 2M RNA expression levels after cel ls were subjected to heat shock. Western blot analysis with 2M monospe cific antibodies recognized a 70-kDa protein which was present in memb rane particles and cytosolic fractions.