6533b7d5fe1ef96bd1263e89

RESEARCH PRODUCT

Cloning of a DNA fragment encoding part of a 70-kDa heat shock protein ofCandida albicans

Rafael SentandreuM. Victoria ElorzaPilar ErolesMaria Sentandreu

subject

Molecular Sequence DataMicrobiologyWestern blotImmunoscreeningHeat shock proteinComplementary DNACandida albicansGeneticsmedicineHSP70 Heat-Shock ProteinsAmino Acid SequenceRNA MessengerCloning MolecularHeat shockDNA FungalCandida albicansMolecular BiologySouthern blotBase Sequencebiologymedicine.diagnostic_testChromosome MappingSequence Analysis DNAbiology.organism_classificationMolecular biologyBiochemistryPolyclonal antibodiesbiology.protein

description

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

https://doi.org/10.1111/j.1574-6968.1995.tb07506.x