Characterization of a cDNA clone, encoding a 70 kDa heat shock protein from the dermatophyte pathogen Trichophyton rubrum.
Trichophyton rubrum is an anthropophilic fungus causing up to 90% of chronic cases of dermatophytosis. To characterize T. rubrum proteins at the molecular level, we established a cDNA library of this pathogen. Here we describe a recombinant cDNA clone identical to eukaryotic 70kDa heat-shock proteins (HSPs). Western blot analysis using an anti HSP70 monoclonal antibody detected a recombinant fusion protein in Escherichia coli transformed with the expression vector containing the cloned cDNA insert. Southern blot analysis of T. rubrum genomic DNA detected no other members of the HSP70 gene family. Further analysis revealed the presence of two introns within the ORF of the HSP70 gene. In Northern blot analysis, the cDNA clone was hybridized to a RNA species of about 3.5kb which was constitutively expressed by cells cultured at 27 degrees C and was strongly up-regulated after culture at 37 degrees C. In summary, we have cloned the first member of the HSP family of dermatophytes and characterized it as a member of the Dnak subfamily of 70kDa HSPs.
Rezaie
S
S
Division of Immunology, Allergy and Infectious Diseases, Department of Dermatology, University of Vienna Medical School, Währinger Gürtel 18-20, A-1090, Vienna, Austria.
Ban
J
J
Mildner
M
M
Poitschek
C
C
Brna
C
C
Tschachler
E
E
D000595:Amino Acid Sequence; D003883:Arthrodermataceae / Q000235:genetics*; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004271:DNA, Fungal; D015723:Gene Library; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics*; D007438:Introns; D008957:Models, Genetic; D008969:Molecular Sequence Data; D012331:RNA, Fungal; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D014249:Trichophyton / Q000235:genetics*
D018076:DNA, Complementary; D004271:DNA, Fungal; D018840:HSP70 Heat-Shock Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D011994:Recombinant Proteins