Transcriptomic/proteomic identification of allergens in the mite Tyrophagus putrescentiae

Abstract

While a number of allergens from house dust mites have been described, much remains to be discovered about allergens from storage mites. Here, next-generation sequencing was combined with MS/MS shotgun proteomics to identify proteins, especially potential allergens from Tyrophagus putrescentiae, commonly found in stored food products, especially flour. cDNAs of suspected allergens were cloned and expressed from bacterial cells, and recombinant allergens were tested for binding to IgE in sera from T. putrescentiae-sensitive patients. These analyses identified three previously uncharacterized allergens, Tyr p 28, Tyr p 35, and Tyr p 36, which have been officially assigned by the WHO/IUIS Allergen Nomenclature Sub-committee. Recombinant proteins rTyr p 28, rTyr p 35, and rTyr p 36 bound with 47.1%, 82.4%, and 70.6% of T. putrescentiae-sensitive sera. We provide here a new method to identify allergens by the combination of transcriptomic and proteomic approaches.

Expression, epitope prediction and IgE‑binding of the <em>Tyrophagus putrescentiae</em> group 13 allergen

“…A series of studies on the T. putrescentiae allergens from China have thus been conducted. Using molecular cloning techniques, the sequencing and characterization of group 4, 28, 35 and 36 T. putrescentiae allergens from China were reported in our previous studies (12,13). The T. putrescentiae group 13 allergen (Tyr p 13) was another allergic component in our series of studies.…”

Transcriptomics-Based Identification of Aquaporin Diversity in the House Dust Mite Dermatophagoides farinae (Acariformes: Pyroglyphidae)

“…As described in our previous study ( Cui et al 2016a ), a transcriptome library was constructed with a SMART cDNA Library Construction Kit (CLONTECH Corporation, Mountain View, CA; Code No. 634901).…”

Identification of three aquaporin subgroups from Blomia tropicalis by transcriptomics

“…Transcriptome library construction, sequencing, de novo assembly and annotation. Previously described methods were used to sequence and assemble the transcriptome of B. tropicalis (15). Briefly, to create a pooled cDNA library for large-scale sequencing, a SMART™ cdNA Library Construction kit (catalog no.…”

“…The use of transcriptomics approaches opens novel possibilities for advancing our understanding of insect genomes (14). In our recent study, RNA-seq, combined with tandem mass spectroscopy, was used to identify and study potential allergens from Tyrogphagus putrescentiae (15), a mite species with importance for human allergic disease.…”