Open AccessThe Complete cDNA Sequence and Expression of the First Major Allergenic Protein of Malassezia Furfur , Mal f 1
Author(s) -
Schmidt Margit,
Zargari Arezou,
Holt Palle,
Lindbom Lars,
Hellman Ulf,
Whitley Paul,
Ploeg Ingeborg,
Härfast Bengt,
Scheynius Annika
Publication year - 1997
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1997.00181.x
Subject(s) - complementary dna , peptide sequence , biology , microbiology and biotechnology , fusion protein , open reading frame , amino acid , nucleic acid sequence , protein sequencing , maltose binding protein , recombinant dna , biochemistry , gene
For the first time the complete cDNA encoding a major allergen and novel protein of the yeast Malassezia furfur , Mai f 1, has been sequenced and expressed. The amino acid sequences of nine tryptic peptides of the protein were determined. Oligonucleotides were designed from these amino acid sequences. The cDNA sequence was obtained by hybridizing these primers to mRNA and enhancement by reverse‐transcriptase PCR techniques. The cDNA is 1176 bp in length. It shows an open reading frame of 1050 bp coding for a protein of 38178 Da and a deduced amino acid sequence containing 350 residues. The hydropathy plot and the tryptic digest indicate that the first 22 amino acids represent a leader sequence determining a mature protein of 35988 Da. The complete encoding cDNA was expressed as a maltose‐binding protein fusion protein in Escherichia coli. The recombinant fusion protein reacted with our specific monoclonal antibody and with IgE from patients with atopic dermatitis.