6533b836fe1ef96bd12a129d

RESEARCH PRODUCT

Cloning of cDNAs coding forCandida albicanscell surface proteins

M V ElorzaMaria SentandreuEulogio ValentínDaniel GozalboRafael Sentandreu

subject

biologycDNA libraryGeneral MedicineMolecular cloningbiology.organism_classificationMolecular biologyInfectious DiseasesPolyclonal antibodiesComplementary DNAImmunoscreeningbiology.proteinGenomic libraryCandida albicansSouthern blot

description

Two cDNA libraries were constructed from mRNAs obtained from yeast cells and germ-tubes of Candida albicans in lambda gt11. Immunoscreening with polyclonal antibodies raised against cell wall components allowed the detection of 29 positive clones. Two of these clones were selected for their specific reactivity with antisera either from yeast (clone 11Y) or germ-tubes (clone 24M). cDNA fragments were isolated by the digestion of lambda DNA with EcoRI. Southern blot analysis with these fragments as probes demonstrated homology with C. albicans DNA, and by Northern analysis two mRNAs transcripts were detected with sizes of approximately 1·5 kb for 11Y and 1·1 kb for 24M. Both transcripts were present in yeast cells as well as in germ-tubes. The whole genes were isolated from a C. albicans genomic library in the YRp7 vector by hybridization with the cDNA probes. Monospecific antibodies were purified from polyclonal antisera by affinity for the fusion proteins. Western blot analysis with 11Y-specific antibodie...

https://doi.org/10.1080/02681219580000231