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Section 4. Medical science
https://doi.org/10.29013/ESR-20-7.8-23-25
Venger Andrii, PhD (biology), associate professor, Odessa national medical university, Ukraine E-mail: venger87@ukr.net Venger Olga,
PhD (biology), junior scientific researcher, Plant breeding and genetics Institute - National center of seed and cultivar investigation, Ukraine
Vasina Yuliia,
student, Odessa national medical university, Ukraine
Zhuravlov Ivan,
lecturer, The state institution South Ukrainian National Pedagogical University named after K. D. Ushynsky, Ukraine
Kholodnitskiy Igor, lecturer, The state institution South Ukrainian National Pedagogical University named after K. D. Ushynsky, Ukraine
Liubov Bahrii,
lecturer, The state institution South Ukrainian National Pedagogical University named after K. D. Ushynsky, Ukraine
POLYMORPHISM OF AMB A I ENCODING GENE OF AMBROSIA ARTEMISIIFOLIA
Abstract. Diagnostics of polymorphism of Amb a I encoding gene of Ambrosia artemisiifolia was conducted. Polymorphism of Amb a I encoding gene of Ukrainian and world collection samples of A. artemisiifolia was researched.
Keywords: Ambrosia artemisiifolia, Amb a I, allergen, polymorphism. Ukrainian samples.
Allergens released by Ambrosia from the Astera- a 26-kDa a-chain and an associated 12-kDa ^-chain
ceae family are an important factor of allergy. Amb [3, 27].
a 1 is the major allergen of Ambrosia. This protein A. artemisiifolia is an invasive weed from North
belongs to the pectate lyase family. Amb a 1 is an America, which has currently inhabited many re-
acidic non-glycosylated 38-kDa protein consisting of gions in Asia and Europe, especially in Ukraine. The
Section 4. Medical science
weed has recently become spreading as a neophyte in Europe, while climate change may also have affected the growth of the plant and additionally may also influence pollen allergenicity [4, 176]. In Ukraine, the number of diseases caused by this allergen has recently increased [1, 350].
The gene for Amb a I had recently been cloned, and it was shown that Amb a I makes a group of proteins with three polymorphic isoform members [2].
Polymorphism of Amb a I encoding gene of Ukrainian samples of A. artemisiifolia is still unknown. The aim of current scientific work was to create primers for polymorphic regions of Amb a I encoding gene and to detect polymorphism of that gene in Ukrainian and world collection samples.
Material and methods
Polymorphism of Amb a I encoding gene of A. artemisiifolia was researched by polymerase chain reaction (PCR) in silico and in vitro. Primers and time-temperature environment for PCR were conducted by VectorNTI10 program.
Table 1. - Products of PCR-
124 nucleotide sequences of Amb a I encoding gene from National Centre of Biotechnology Information and 43 DNA samples of Ukrainian A. artemisiifolia were analysed by PCR in silico with Fast-PCR program [5].
Polymorphism of Amb a I encoding gene in DNA, extracted from 43 samples Ukrainian A. artemisiifolia was analysed by PCR in vitro. Sizes of products of PCR were detected by gel-electrophoresis.
Results
By VectorNTI10 program for analysis of polymorphism of Amb a I encoding gene of A. artemisiifolia there were conducted following primers (5'->3'): AMB1F GTCTACACGGTCACCAG-CAA and AMB1R AGGGGCC AAG AATCT-GTTCC. Condition of PCR: 2 min at 94 0 C for first denaturation; 35 basic cycles of 0,5 min at 94 oC, 1 min at 54 oC, 1,5 min at 72 oC; and 10 min at 72 oC for final elongation.
Results of PCR are present in (table 1).
nalysis in silico and in vitro
Type of PCR Products of PCR, base pairs
690.690 690.700 700.700
Number of samples
in silico 88 36
in vitro 27 12 4
Discussion
The most popular combination of products of amplification in silico and in vitro is 690.690 base pairs (bp). Combination 690.700 bp was also obtained after both providing types of PCR and combination 700.700 bp was produced only in reaction with DNA of Ukrainian samples of A. artemisiifolia. Unique combination of amplicons can be correlated with unique germplasm presence in Ukrainian population of A. artemisiifolia.
Conclusion
In result of the accomplished research the primers and conditions of PCR for detection of polymorphism of Amb a I encoding gene of A. artemisiifolia were conducted. Polymorphism of Amb a I encoding gene with nucleotide sequences from NCBI and DNA of Ukrainian samples of A. artemisiifolia was researched. Unique combination of alleles in Ukrainian population of A. artemisiifolia was described.
References:
1. Chen M., Xu J. D. Devis, J.n Shi K. Ren I. Searle D. Zhang. Origin and Functional Prediction of Pollen Allergens in Plants. Plant Physiol. 172(1). 2016.- P. 341-357.
2. Chruszcz M., Kapingidza A. B., Dolamore C., Kowal K. A robust method for the estimation and visualization of IgE cross-reactivity likelihood between allergens belonging to the same protein family. PLoS One. 13(11). 2018. e0208276.
3. Hiller K. M., Lubahn B. C., Klapper D. G. Cloning and expression of ragweed allergen amb a 6. Scandinavian Journal of Immunology, 48. 1998.- P. 26-36.
4. Kelish A. L., Zhao F., Heller W., Durner J., Winkler J. B., Behrendt H., Traidl-Hoffmann C., Horres R., Pfeifer M., Frank U. Ragweed (Ambrosia artemisiifolia) pollen allergenicity: SuperSAGE transcriptomic analysis upon elevated CO2 and drought stress. BMC Plant Biology, 14. 2014.- 176 p. Published online.
5. URL: http://www.ncbi.nlm.nih.gov
