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2 2021

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Utilization of specific primers in legume allergens based polymorphism screening

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Klongová L. , Kováčik A. , Urbanová L. , Kyseľ M. , Ivanišová E. , Žiarovská J.

Science, Technology and Innovation

2021

tom Vol. 13, no. 2

12--21

Different types of allergies became a part of life of many people around the world. The research activities connecting to allergens are actually not oriented only for protein and immunological interactions, but to the genomic and transcriptomic background of them, too. Analysis and description of genomic variability of allergens in plant food resources will help to manage the allergen based strategies in the future. Here, the bioinformatic approach was used to develop and validate the specific primers for genomic screening of polymorphism of profilins (Profilin Based Amplicon Polymorphism; PBAP) and vicilins (Vicilin Based Amplicon Polymorphism; VBAP) among the legumes. The alignment of existing public databases data for these allergens in the group of legumes was performed. Subsequently, specific primers were designed and their ability to generate polymorphic amplicons were tested for three legumes – bean, lentil and chickpeas. In all cases, amplicons were generated and polymorphism was detected in all three species for profilin as well as for vicilin.

BBAP amplification profiles of apple varieties

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Speváková I. , Urbanová L. , Kyseľ M. , Bilčíková J. , Farkasová S. , Žiarovská J.

Science, Technology and Innovation

2021

tom Vol. 13, no. 2

1--6

Several types of allergies are currently known and are characterized by an exaggerated response of the immune system to substances from various sources called allergens. One of them is a food allergy, which is becoming more common in the population. For this reason, it is necessary to describe the issue from several aspects including genomic variability of plant allergens. The objective of this study was to analyse intraspecific variability of Bet v 1 of 10 different varieties of apple species (Malus domestica Borkh.). BBAP technique for genomic determination of the presence of Bet v 1 homologs at the DNA level was performed. Degenerate primers that anneal a variable and conserved part of PR-10 protein homologues genes were used in the analyse. Amplicons were generated and formed relatively monomorphic profiles, indicating the stability of the given isoforms of Bet v 1 within the selected apple varieties. To evaluate the potential allergenicity of selected varieties further studies on another molecular level such as a comparison of gene expression of the PR-10 family members and their protein expression levels are needed.