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Assessment of DNA Contamination in RNA Samples Based on Ribosomal DNA
Published on: January 22, 2018
Nucleotide sequence analysis of a complementary DNA coding for a Blomia tropicalis allergen
L Puerta1, L Caraballo, E Fernández-Caldas
1Institute of Immunological Research, University of Cartagena, Colombia.
The Journal of Allergy and Clinical Immunology
|November 1, 1996
Summary
Researchers identified a new major allergen from the Blomia tropicalis mite, important in tropical allergies. This recombinant protein, Bt11a, was recognized by IgE in over half of tested asthma patients, indicating its allergenic significance.
Area of Science:
- Immunology
- Molecular Biology
- Allergology
Background:
- Blomia tropicalis mites are significant allergens in tropical and subtropical regions.
- Understanding mite allergens is crucial for diagnosing and treating allergic diseases like asthma.
Purpose of the Study:
- To identify and characterize a novel allergen from Blomia tropicalis.
- To assess the allergenic potential of the identified recombinant protein.
Main Methods:
- Construction and expression of a Blomia tropicalis complementary DNA (cDNA) clone (Bt11a) in lambda phage.
- Analysis using plaque radioimmunoassay and IgE binding assays with patient sera.
- Sequencing of the cDNA insert using polymerase chain reaction (PCR) and various primers.
Main Results:
- The recombinant allergen (Bt11a) was recognized by IgE in 16 out of 32 sera from asthma patients with a positive RAST response.
- The full cDNA sequence revealed a 432 bp reading frame encoding a mature protein of 124 residues (14,206 Da) with a putative signal peptide.
- No homology to known sequences or N-linked glycosylation sites were found in the nucleotide or amino acid sequences.
Conclusions:
- The recombinant protein derived from the Bt11a clone represents a significant major allergen of the Blomia tropicalis mite.
- This finding contributes to the understanding of Blomia tropicalis-induced allergies and may aid in diagnostic and therapeutic strategies.
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