Nucleotide sequence analysis of a complementary DNA coding for a Blomia tropicalis allergen.
Blomia tropicalis is a mite of allergenic importance in tropical and subtropical areas. A clone (Bt11a) from a B. tropicalis complementary DNA library was expressed in lambda phage and analyzed by plaque radioimmunoassay. The recombinant allergen produced by this clone was bound by IgE in 16 of 32 sera from individuals with asthma with a positive RAST response and none of 3 control sera from healthy individuals with negative RAST response to B. tropicalis. The cDNA insert was amplified by polymerase chain reaction with use of universal primers. A 582-base-pair (bp) fragment was cloned into a pCR II vector. The complete sequence of both strands was determined by using T7, SP6, and internal primers. The sequence shows a 432 bp reading frame with a 34 bp 5' untranslated region and a 116 bp 3' untranslated region with a poly A tail. Analysis of the sequence suggests that it encodes a putative signal peptide of 20 residues and a 124-residue mature protein allergen of 14,206 Da. The nucleotide and the inferred amino acid sequences did not show homology to any known sequence. No potential N-linked glycosylation site was found. The recombinant protein appears to represent a major allergen of the mite B. tropicalis.
Puerta
L
L
Institute of Immunological Research, University of Cartagena, Colombia.
Caraballo
L
L
Fernández-Caldas
E
E
Avjioglu
A
A
Marsh
D G
DG
Lockey
R F
RF
Dao
M L
ML
D000328:Adult; D000485:Allergens / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001249:Asthma / Q000235:genetics*; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics*; D006801:Humans; D007073:Immunoglobulin E / Q000096:biosynthesis / Q000276:immunology*; D008925:Mites / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011852:Radioallergosorbent Test; D011994:Recombinant Proteins / Q000235:genetics; D012882:Skin Tests
D000485:Allergens; D018076:DNA, Complementary; D011994:Recombinant Proteins; D007073:Immunoglobulin E