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EN
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.
EN
In our experiment, 3 species-specific primer pairs cultivated in cell lines were used: Encephalitozoon cuniculi-specific primer pairs (ECUNF and ECUNR), Encephalitozoon hellem-specific primer pairs (EHELF and EHELR), and Encephalitozoon intestinalis-specific primer pairs (SINTF and SINTR). The PCR products were estimated to be 550 bp in E. cuniculi, 547 bp in E. hellem and 545 bp in E. intestinalis respectively, which can prove the precision and reliability of this method in the species identification of the genus Encephalitozoon. All 3 primer pairs were species-specific and none of them amplified gene sequences from other Encephalitozoon spp.
EN
Both native European species, Bursaphelenchus mucronatus and B. fraudulentus, are relatively common and harmless to trees. They belong to the xylophilus group, which includes also the quarantine pest B. xylophilus – the causal agent of the pine wilt disease. They can be difficult to distinguish morphologically from each other and from B. xylophilus. Therefore, reliable methods of taxonomic identification of these species are therefore of particular interest to plant quarantine services. PCR amplification with specific primers enables rapid and precise species identification, even from a single nematode. In 2004, Matsunaga and Togashi designed specific primers for B. xylophilus and B. mucronatus. The aim of this study was to design specific primers for B. fraudulentus. The specificity of the newly designed primers was tested on eight isolates of B. fraudulentus which originated from different parts in Europe (Austria, Germany, Poland, Russia and Ungarn). For all isolates, PCR amplification resulted in products of identical length of 617 bp, but no PCR products were generated for B. xylophilus and B. mucronatus. The PCR amplification with primer sets specific for B. xylophilus (XF and XR), B. mucronatus (MF and MR) and those for B. fraudulentus (FF and FR) designed in this study resulted in amplicons of different lengths (557, 210 and 617 bp, respectively), which can be easily distinguished in agarose gels.
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