Open AccessCloning and characterisation of a group II allergen from the dust mite Tyrophagus putrescentiae
Author(s) -
Eriksson Tove Lisa Jenny,
Johansson Eva,
Whitley Paul,
Schmidt Margit,
Elsayed Said,
van HageHamsten Marianne
Publication year - 1998
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1327.1998.2510443.x
Subject(s) - tyrophagus putrescentiae , complementary dna , biology , peptide sequence , nucleic acid sequence , house dust mite , primer (cosmetics) , allergen , microbiology and biotechnology , genetics , amino acid , mite , gene , antibody , chemistry , allergy , immunoglobulin e , botany , organic chemistry , immunology
The complete cDNA encoding a major allergen from the dust mite Tyrophagus putrescentiae , Tyr p 2, has been sequenced and expressed. A degenerate primer was designed to the N‐terminal amino acid sequence of the 16‐kDa protein. The complete cDNA sequence was achieved by using reverse transcriptase PCR, PCR+1, standard cloning and sequencing techniques. The cDNA of Tyr p 2 is 552 nucleotides in length from the start codon including 126 nucleotides after the stop codon up to the beginning of the poly(A) tail. The leader sequence consists of 15 amino acids. Regarding the predicted amino acid sequence, there are no potential N‐glycosylation sites (N‐X‐S/T). The sequence showed similarity to group II allergens from other mite species, and some regions are completely conserved. To show that the cloned cDNA sequence was coding for an allergen, Tyr p 2 was expressed in Escherichia coli and shown to react with a T. putrescentiae ‐positive serum pool.