Open AccessCloning of a DNA fragment encoding part of a 70‐kDa heat shock protein of Candida albicans
Author(s) -
Eroles Pilar,
Sentandreu Maria,
Elorza M. Victoria,
Sentandreu Rafael
Publication year - 1995
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1995.tb07506.x
Subject(s) - immunoscreening , microbiology and biotechnology , complementary dna , biology , candida albicans , polyclonal antibodies , southern blot , open reading frame , northern blot , cdna library , molecular cloning , western blot , sequence analysis , heat shock protein , blot , hspa14 , dna , gene , peptide sequence , antibody , biochemistry , hsp70 , genetics
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.