EPHA1 gene SNPs analysis in population of Ukraine
Aim. Analysis the EPHA1 gene G1475A and G1891A alleles distribution in the population of Ukraine, and to study the protein secondary structure as the first step in the investigation of EPHA1 gene involvement in intellectual disability pathogenesis. Methods. Observation group consisted of 300 individ...
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| Cite this: | EPHA1 gene SNPs analysis in population of Ukraine / R.V. Gulkovskyi, S.Y. Chernushyn, S.A. Kravchenko, G.M. Bychkova, L.A. Livshits // Вiopolymers and Cell. — 2013. — Т. 29, №. 6. — С. 506-510. — Бібліогр.: 15 назв. — англ. |
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nasplib_isofts_kiev_ua-123456789-1536752025-02-09T21:51:15Z EPHA1 gene SNPs analysis in population of Ukraine Аналіз SNPs гена EPHA1 у популяції України Анализ SNPs гена EPHA1 в популяции Украины Gulkovskyi, R.V. Chernushyn, S.Y. Kravchenko, S.A. Bychkova, G.M. Livshits, L.A. Biomedicine Aim. Analysis the EPHA1 gene G1475A and G1891A alleles distribution in the population of Ukraine, and to study the protein secondary structure as the first step in the investigation of EPHA1 gene involvement in intellectual disability pathogenesis. Methods. Observation group consisted of 300 individuals, including 164 (54.6 %) male and 136 (45.3 %) female individuals. Polymorphic variants were detected using PCR followed by Kpn1 RFLP analysis for G1475A and ARMS PCR analysis for G1891A. Results. The data concerning EPHA1 genotypes and allelic variants distribution were obtained. The low frequency of 1475A allele (0,012) and the absence of 1891A allele (not previously described) were revealed in the population of Ukrainian. Conclusions. Assays for the detection of EPHA1 gene G1475A and G1891A SNPs based on ARMS and restriction analysis were developed and the preliminary data on distribution of G1475A and G1891A polymorphisms in the population of Ukraine were obtained. Мета. Аналіз розподілу алельних варіантів поліморфізмів G1475A і G1891A гена EPHA1 у популяції України та вивчення вторинної структури білка як перший етап визначення ролі гена EPHA1 у патогенезі інтелектуальної недієздатності. Методи. Алельні варіанти SNP для G1475A і G1891A досліджували методами ПЛР- ПДРФ та алель-специфічної ПЛР відповідно. Результати. Отримано дані стосовно розподілу генотипів і алельних варіантів гена EPHA1. Виявлено низьку частоту алеля 1475A (0,012) та відсутність алеля 1891A в популяції України. Висновки. Створено методики детекції замін G1475A і G1891A в гені EPHA1 та проведено попередні дослідження розподілу поліморфізмів G1475A і G1891A у популяції Україні. Цель. Анализ распределения аллельных вариантов полиморфизмов G1475A и G1891A гена EPHA1 в популяции Украины и изучение вторичной структуры белка как первый этап определения роли гена EPHA1 в патогенезе интеллектуальной недееспособности. Методы. Аллельные варианты SNP для G1475A и G1891A исследовали методами ПЦР-ПДРФ и аллель-специфической ПЦР соотетственно. Результаты. Получены данные относительно распределения генотипов и аллельных вариантов гена EPHA1. Выявлена низкая частота аллеля 1475A (0,012) и отсутствие аллеля 1891A в популяции Украины. Выводы. Созданы методики обнаружения замен G1475A и G1891A в гене EPHA1 и проведены предварительные исследования распределения полиморфизмов G1475A и G1891A в популяции Украины. 2013 Article EPHA1 gene SNPs analysis in population of Ukraine / R.V. Gulkovskyi, S.Y. Chernushyn, S.A. Kravchenko, G.M. Bychkova, L.A. Livshits // Вiopolymers and Cell. — 2013. — Т. 29, №. 6. — С. 506-510. — Бібліогр.: 15 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.000841 https://nasplib.isofts.kiev.ua/handle/123456789/153675 577.11 + 577.21 en Вiopolymers and Cell application/pdf Інститут молекулярної біології і генетики НАН України |
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Biomedicine Biomedicine Gulkovskyi, R.V. Chernushyn, S.Y. Kravchenko, S.A. Bychkova, G.M. Livshits, L.A. EPHA1 gene SNPs analysis in population of Ukraine Вiopolymers and Cell |
| description |
Aim. Analysis the EPHA1 gene G1475A and G1891A alleles distribution in the population of Ukraine, and to study the protein secondary structure as the first step in the investigation of EPHA1 gene involvement in intellectual disability pathogenesis. Methods. Observation group consisted of 300 individuals, including 164 (54.6 %) male and 136 (45.3 %) female individuals. Polymorphic variants were detected using PCR followed by Kpn1 RFLP analysis for G1475A and ARMS PCR analysis for G1891A. Results. The data concerning EPHA1 genotypes and allelic variants distribution were obtained. The low frequency of 1475A allele (0,012) and the absence of 1891A allele (not previously described) were revealed in the population of Ukrainian. Conclusions. Assays for the detection of EPHA1 gene G1475A and G1891A SNPs based on ARMS and restriction analysis were developed and the preliminary data on distribution of G1475A and G1891A polymorphisms in the population of Ukraine were obtained. |
| format |
Article |
| author |
Gulkovskyi, R.V. Chernushyn, S.Y. Kravchenko, S.A. Bychkova, G.M. Livshits, L.A. |
| author_facet |
Gulkovskyi, R.V. Chernushyn, S.Y. Kravchenko, S.A. Bychkova, G.M. Livshits, L.A. |
| author_sort |
Gulkovskyi, R.V. |
| title |
EPHA1 gene SNPs analysis in population of Ukraine |
| title_short |
EPHA1 gene SNPs analysis in population of Ukraine |
| title_full |
EPHA1 gene SNPs analysis in population of Ukraine |
| title_fullStr |
EPHA1 gene SNPs analysis in population of Ukraine |
| title_full_unstemmed |
EPHA1 gene SNPs analysis in population of Ukraine |
| title_sort |
epha1 gene snps analysis in population of ukraine |
| publisher |
Інститут молекулярної біології і генетики НАН України |
| publishDate |
2013 |
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Biomedicine |
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https://nasplib.isofts.kiev.ua/handle/123456789/153675 |
| citation_txt |
EPHA1 gene SNPs analysis in population of Ukraine / R.V. Gulkovskyi, S.Y. Chernushyn, S.A. Kravchenko, G.M. Bychkova, L.A. Livshits // Вiopolymers and Cell. — 2013. — Т. 29, №. 6. — С. 506-510. — Бібліогр.: 15 назв. — англ. |
| series |
Вiopolymers and Cell |
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| fulltext |
506
UDC 577.11 + 577.21
EPHA1 gene SNPs analysis in population of Ukraine
R. V. Gulkovskyi1, 2, S. Y. Chernushyn1, S. A. Kravchenko1,
G. M. Bychkova1, L. A. Livshits1
1Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
2Taras Shevchenko National University of Kyiv, Educational and Scientific Centre «Institute of Biology»
64, Volodymyrska Str., Kyiv, Ukraine, 03601
livshits@imbg.org.ua
Aim. Analysis the EPHA1 gene G1475A and G1891A alleles distribution in the population of Ukraine, and to
study the protein secondary structure as the first step in the investigation of EPHA1 gene involvement in
intellectual disability pathogenesis. Methods. Observation group consisted of 300 individuals, including 164
(54.6 %) male and 136 (45.3 %) female individuals. Polymorphic variants were detected using PCR followed by
Kpn1 RFLP analysis for G1475A and ARMS PCR analysis for G1891A. Results. The data concerning EPHA1
genotypes and allelic variants distribution were obtained. The low frequency of 1475A allele (0,012) and the
absence of 1891A allele (not previously described) were revealed in the population of Ukrainian. Conclusions.
Assays for the detection of EPHA1 gene G1475A and G1891A SNPs based on ARMS and restriction analysis were
developed and the preliminary data on distribution of G1475A and G1891A polymorphisms in the population of
Ukraine were obtained.
Keywords: EPHA1 gene, polymorphism, population.
Introduction. In the frame of CHERISH project (no.
223692) concerning the intellectual disability (ID) ge-
netic basis, the next generation exome sequencing was
conducted for a Ukrainian family (UKR 094) with two
affected children (proband and his younger sibling) and
their healthy non-consanguineous parents. The proband
(UKR 263) is a 12 year old boy with ID (IQ = 43) and
hyperactivity. His brother (UKR 264) is a 4 year old boy
with febrile convulsions, ID and multiple congenital ano-
malies (Fig. 1). Both affected probands underwent phy-
sical and neurological examination, standard G-ban-
ding karyotype analysis and biochemical tests: amino-
acids and acylcarnitines (TANDEM MS) as well as mo-
lecular genetic screening of common ID syndromes
(FRAXA, FRAXE, FRAXF loci together with 15q11-
q13 locus). Standard karyotype, biochemical and mo-
lecular genetic tests as well as array-CGH analysis (400
K resolution) were normal in both patients.
Exome analysis revealed several variants in either
homozygous or compound heterozygous state in five
genes. Among these genes, we decided to concentrate
on the EPHA1 gene, where two non-synonymous sub-
stitutions were detected in both patients: G1475A
(rs11768549) inherited from the mother and G1891A
(has not been previously described) inherited from the
father. The both non-synonymous SNPs have been
identified in the coding region (exons 7 and 11) of the
EPHA1 receptor gene. EPHA1 has been reported to be
highly expressed in the neural tissues during embryo-
genesis and it seemed a good candidate for autosomal
recessive inherited ID [1, 2].
EPHA1 is the founding member of the erythropo-
ietin-producing hepatocellular (Eph) receptors family,
but little is known about its function. Eph kinases con-
stitute the largest family of receptor tyrosine kinases,
with 16 distinct members. The receptors are termed
EPHA (EPHA1–8, only including those found in mam-
mals) or EPHB (EPHB1–4, EPHB6) on the basis of se-
ISSN 0233–7657. Biopolymers and Cell. 2013. Vol. 29. N 6. P. 506–510 doi: 10.7124/bc.000841
� Institute of Molecular Biology and Genetics, NAS of Ukraine, 2013
quence homologies and depending on the subgroup of
ligands that they bind [3]. Ligand for the EPHA recep-
tor is called ephrin-A. It has been recently demonstrated
that EPHA/ephrin-A signalling has a proapoptotic effect
on the embryonic cortex without affecting the prolife-
ration or cell cycle progression [4]. Ephrin and Eph re-
ceptor genes are implicated in the control of cell and
axon guidance in neural systems of different species [1,
2, 5, 6].
EPHA1 gene is located at chromosome 7q34 and
contains 18 exons, two more than the related tyrosine
kinase. The human EPHA1 gene is transcribed as a sing-
le 3.5 kb mRNA, and encodes a polypeptide of 984 ami-
no acids with a molecular mass of 130 kDa [7]. It is im-
portant to know that EPHA1 is characterized by high
conservative level among many species, indicating that
there may be an evolutionarily conserved function [7].
It was shown that this gene is highly expressed in liver,
lung, and kidney, but not in the adult brain. However, it
is highly expressed in the spinal neural tube during neu-
rulation [8]. The role of EPHA1 during tissue morpho-
genesis is highlighted by the expression studies perfor-
med on mouse embryos, where the pattern of expression
is dynamic throughout the immediate postimplantation,
gastrulation, neurulation, and early somitogenesis pe-
riods [9].
In this study the analysis of single nucleotide poly-
morphisms G1475A and G1891A in EPHA1 gene was
performed in population of Ukraine and it is the first
step in the analysis of EPHA1 gene involvement in in-
tellectual disability pathogenesis. To understand a bio-
chemical effect of these substitutions we tried to predict
possible changes in the EPHA1 protein structure.
Materials and methods. The DNA-samples were
extracted from peripheral blood leucocytes of unrela-
ted volunteer donors from different regions of Ukraine
by standard methods [10]. Informed consents were ob-
tained from all individuals participating in our study.
The observation group consisted of 300 individuals,
including 164 (54.6 %) male and 136 (45.3 %) female
individuals. This group may be considered representati-
ve for the estimation of DNA polymorphism frequency
in autosomal genes [11, 12].
DNA sequencing was performed by the dideoxy-
nucleotide chain-termination method [13] using [35S]-
dATP or [35S]-dCTP (ICN), the Sequenase version 2.0
DNA sequencing kit (USB) and Thermo Sequenase cyc-
le sequencing kit («Amersham», Great Britain).
An analysis of the G1891A polymorphism was per-
formed by ARMS (amplification refractory mutation
system) PCR. Specific oligonucleotide primers were
complementary either to the wild type or to the sequen-
ce with substitution (Table 1). The PCR amplification
was performed in a final volume of 15 �l containing 1�
PCR buffer, 1.5 mM MgCl2, 200 �M of each dNTP,
0.8 �M of each primer, 0.2 units of Taq-DNA polyme-
rase and 200 ng of the DNA template. The cycling con-
ditions for G1891A variant were as follows: initial de-
naturation at 94 oC for 5 min, 30 cycles consisting of de-
naturation at 94 oC for 30 s, annealing at 63 oC for 30 s,
extension at 72 oC for 30 s, and a final elongation step at
72 oC for 5 min.
The presence of G1475A EPHA1 polymorphism
was examined by PCR-RFLP (restriction fragment length
polymorphism) analysis using the specific oligonucleo-
tide primers described in the table. The PCR amplifica-
507
EPHA1 GENE SNPs ANALYSIS IN POPULATION OF UKRAINE
I
II
III
1 2 3 4 5 6
1 2 3 4 5
1 2 3 4
Fig. 1. Family UKR 094 pedigree: III-4 – UKR 263; III-5 – UKR 264;
II-3 – UKR 265; II-4 – UKR 266
Substitution Nucleotide sequence
Amplicon
size, bp
G1475A
AAAGGGCCAGGACCCAGTGGGGTC
AGC – forward
194
CATGTGCTCTGATGCTGTCC –
reverse
–
G1891A
CCTCTTTGAACCATTGCGTT –
common
204
GCTGATGGTGGACACTGTCATAG –
wild type
–
GCTGATGGTGGACACTGTCATAA –
polymorphic
–
Table 1
Specific oligonucleotides, designed and synthesized in accordance to
corresponding exon sequences of EPHA1 gene, were used as primers
tion was performed in a final volume of 25 �l contai-
ning 1 � PCR buffer, 1.5 mM MgCl2, 200 �M of each
dNTP, 1 �M of each primer, 0.2 units of Taq-DNA po-
lymerase and 200 ng of the DNA template. The cycling
conditions for G1475A variant were as follows: initial
denaturation at 94 oC for 5 min, 30 cycles consisting of
denaturation at 94 oC for 40 s, annealing at 55 oC for
40 s, extension at 72 oC for 40 s, and a final elongation
step at 72 oC for 3 min. The amplified fragments were
digested with KpnI. Digestion was performed in 15 �l
reaction volume containing 1 X reaction buffer, 0.5 units
of the restriction enzyme and 10 �l of purified PCR
product, incubated at 37 oC overnight and analyzed by
6 % standard agarose gel electrophoresis.
Primers were designed using the web-based PRIMER
3.0 program (http://workbench.sdsc.edu). We used the
«BLAST» program at http://www.ncbi.nlm.nih.gov/
blast to check the primers specificity. Hypothetical RFLP
results were tested using NEBcutter V2.0 (http://tools.
neb.com/NEBcutter2). We also used the PSIPRED Pro-
tein Sequence Analysis Workbench (http://bioinf.cs.ucl.
ac.uk/psipred) to predict the protein secondary structure.
Results and discussion. The Sanger sequencing of
EPHA1 gene loci with substitutions was performed. Se-
quencing confirmed the results of the exome analysis
of UKR 094 family members and revealed that the fa-
ther is a heterozygous carrier for the C1891T polymor-
phism and the mother is a heterozygous carrier for the
C1475T polymorphism (Fig. 2). The sample that had
previously undergone Sanger sequencing was used as a
positive control.
RFLP analysis of the G1475A EPHA1 variants. The
designed primers successfully amplified the exon 7 frag-
ment (194 bp) of EPHA1 gene. The G to A transition in
G1475A variant creates a restriction site for endonuc-
lease KpnI. Thereby three different patterns could be
observed for G1475A variant after the restriction
digestion: a 194 bp band (for 1475A/A); a 194 bp, a
101 bp and a 93 bp bands (for 1475G/A); a 101 bp and a
93 bp bands (1475G/G) (Fig. 3).
ARMS for G1891A analysis is based on the obser-
vation that PCR amplification is inefficient or comple-
tely refractory if there is a mismatch between the 3' ter-
minal nucleotide of a PCR primer and a corresponding
template. This approach implies two PCR reactions.
Amplification of the allele 1891G is accomplished using
a complementary primer «wild type» (Table 1). Con-
versely, only the 1891A allele will be amplified if the 3'
residue is complementary to the sequence with substitu-
tion (Table 1). Thus, as a result a normal (G1891G) in-
dividual generates PCR product only in the «normal»
reaction; a heterozygote gives products in both reac-
tions, and a homozygous A1891A individual gives amp-
lification only in the «mutant» reaction. The results of
analysis of individuals with different genotypes are pre-
sented in Fig. 4.
Based on the RFLP analysis of G1475A variant, indi-
viduals were classified into three groups: GG (1475G/
G), GA (1475G/A) and AA (1475A/A). Of the 300 ana-
lyzed samples from the present study, we have found 7
heterozygous carriers for the G1475A polymorphism
only. Genotypes and allele frequencies of the G1475A
508
GULKOVSKYI R. V. ET AL.
1 2 3 4 5 6 7
� 93
� 101
� 194
bp
Fig. 3. RFLP analysis of G1475A EPHA1 receptor gene variant (4 %
agarose gel electrophoresis): 1 – molecular weight marker (Ladder 50 bp);
2, 5, 6 – individuals with homozygous genotype G1475G; 3, 4 – in-
dividual with heterozygous genotype G1475A; 7 – negative control
A
B
Fig. 2. Representative sequence chromatogram of the EPHA1 gene
exons 7 (A) and 11 (B) PCR products: A – UKR 265 (mother) – hetero-
zygous carrier for the G1475A polymorphism; B – UKR 266 (father) –
heterozygous carrier for the G1891A polymorphism
polymorphism are presented in Table 2. The observed
genotype distributions showed no deviations from Har-
dy-Weinberg expectations.
The frequency of allelic variants of EPHA1 gene in
our study is in agreement with the published data ob-
tained within the International HapMap Project (http://
hapmap.ncbi.nlm.nih.gov/) (Table 2).
We did not identify G1891A substitution in any of
300 investigated volunteer donors. It was found only in
the members of aforesaid Ukrainian family with ID in
children. Previous reports have demonstrated a signifi-
cant association of T3022C (rs11767557) polymor-
phism within this gene with Alzheimer’s disease [14,
15]. EPHA1 has been reported to be highly expressed
in the neural tissues during embryogenesis [3, 4].
The first analyzed SNP – G1475A – exchanges co-
don 492 from arginine (CGG) to a glutamine (CAG).
The second one – G1891A – leads to an amino acid
change at position 631 from glycine (GGA) to arginine
(AGA). To understand a biochemical effect of these
substitutions we tried to predict possible changes in the
EPHA1 protein structure. A comparative analysis of the
predicted secondary structure revealed that both amino
acid substitutions lead to significant changes in the pro-
bability of the �-sheet forming. The change of Arg492
to Glu results in extending the 11th and 12th
�-sheets in
the fibronectin type III repeat of EPHA1 protein by 1
amino acid. The change of Gly631 to Arg results in
extending of the 2nd
�-sheet in the tyrosine kinase do-
main by 1 amino acid. Thereby these substitutions may
directly cause changes in protein solubility or binding
with ligands or protein-partners.
In the frame of CHERISH FP7 project, the exome
NGS was performed for two affected probands from
Ukrainian family with moderate ID, which revealed
two polymorphisms in the EPHA1 gene: G1475A and
novel SNP G1891A. The assays for detection of these
SNPs based on ARMS and restriction analysis were
developed and the preliminary studies on the distribu-
tion of G1475A and G1891A polymorphisms in the
population of Ukraine were conducted. Further case-
control EPHA1 exons sequencing will be carried out to
evaluate the involvement of the EPHA1 mutations in
ID development.
Acknowledgements. We thank M. Noukas, M. Sauk,
L. Milani, T. Pippucci, F. Balombo for their kind assis-
tance in next generation exome sequencing of the Ukr
094 family members. We thank V. Kitam for his kind as-
sistance in analysis of the proteins secondary structure.
Ð. Â. Ãóëêîâñüêèé, Ñ. Þ. ×åðíóøèí, Ñ. À. Êðàâ÷åíêî, Ã. Ì. Áè÷êî-
âà, Ë. À. ˳âøèöü
Àíàë³ç SNPs ãåíà EPHA1 ó ïîïóëÿö³¿ Óêðà¿íè
Ðåçþìå
Ìåòà. Àíàë³ç ðîçïîä³ëó àëåëüíèõ âàð³àíò³â ïîë³ìîðô³çì³â G1475A
³ G1891A ãåíà EPHA1 ó ïîïóëÿö³¿ Óêðà¿íè òà âèâ÷åííÿ âòîðèííî¿
ñòðóêòóðè á³ëêà ÿê ïåðøèé åòàï âèçíà÷åííÿ ðîë³ ãåíà EPHA1 ó ïà-
òîãåíåç³ ³íòåëåêòóàëüíî¿ íå䳺çäàòíîñò³. Ìåòîäè. Àëåëüí³ âàð³-
509
EPHA1 GENE SNPs ANALYSIS IN POPULATION OF UKRAINE
1 2 3 4 5 6 7 8 9
G A G A G A G A
bp
204
Fig. 4. ARMS analysis of G1891A EPHA1 receptor gene variant (2 %
agarose gel electrophoresis): 1 – molecular weight marker (50 bp lad-
der); 2, 3 – individual with heterozygous genotype G1891A; 4–7 – indi-
viduals with homozygous genotype G1891G; 8, 9 – negative control
Populations
Chromosome
sample count
Homozygote GG
(wild type)
Heterozygote GA
Homozygote AA
(polymorphic allele)
Polymorphic allele
frequency
Ukraine 600 0.977 0.023 0.000 0.012
European 226 0.965 0.035 0.000 0.018
Asian 174 1.000 0.000 0.000 0.000
African 116 1.000 0.000 0.000 0.000
Table 2
Genotype and allele frequencies distribution of EPHA1 gene variants in different populations
510
GULKOVSKYI R. V. ET AL.
àíòè SNP äëÿ G1475A ³ G1891A äîñë³äæóâàëè ìåòîäàìè ÏËÐ-
ÏÄÐÔ òà àëåëü-ñïåöèô³÷íî¿ ÏËÐ â³äïîâ³äíî. Ðåçóëüòàòè. Îòðè-
ìàíî äàí³ ñòîñîâíî ðîçïîä³ëó ãåíîòèï³â ³ àëåëüíèõ âàð³àíò³â ãåíà
EPHA1. Âèÿâëåíî íèçüêó ÷àñòîòó àëåëÿ 1475A (0,012) òà â³äñóò-
í³ñòü àëåëÿ 1891A â ïîïóëÿö³¿ Óêðà¿íè. Âèñíîâêè. Ñòâîðåíî ìåòî-
äèêè äåòåêö³¿ çàì³í G1475A ³ G1891A â ãåí³ EPHA1 òà ïðîâåäåíî
ïîïåðåäí³ äîñë³äæåííÿ ðîçïîä³ëó ïîë³ìîðô³çì³â G1475A ³ G1891A
ó ïîïóëÿö³¿ Óêðà¿í³.
Êëþ÷îâ³ ñëîâà: ãåí EPHA1, ïîë³ìîðô³çì, ïîïóëÿö³ÿ.
Ð. Â. Ãóëêîâñêèé, Ñ. Þ. ×åðíóøèí, Ñ. À. Êðàâ÷åíêî,À. Ì. Áû÷êîâà,
Ë. À. Ëèâøèö
Àíàëèç SNPs ãåíà EPHA1 â ïîïóëÿöèè Óêðàèíû
Ðåçþìå
Öåëü. Àíàëèç ðàñïðåäåëåíèÿ àëëåëüíûõ âàðèàíòîâ ïîëèìîðôèçìîâ
G1475A è G1891A ãåíà EPHA1 â ïîïóëÿöèè Óêðàèíû è èçó÷åíèå
âòîðè÷íîé ñòðóêòóðû áåëêà êàê ïåðâûé ýòàï îïðåäåëåíèÿ ðîëè
ãåíà EPHA1 â ïàòîãåíåçå èíòåëëåêòóàëüíîé íåäååñïîñîáíîñòè.
Ìåòîäû. Àëëåëüíûå âàðèàíòû SNP äëÿ G1475A è G1891A èññëå-
äîâàëè ìåòîäàìè ÏÖÐ-ÏÄÐÔ è àëëåëü-ñïåöèôè÷åñêîé ÏÖÐ ñîîò-
âåòñòâåííî. Ðåçóëüòàòû. Ïîëó÷åíû äàííûå îòíîñèòåëüíî ðàñ-
ïðåäåëåíèÿ ãåíîòèïîâ è àëëåëüíûõ âàðèàíòîâ ãåíà EPHA1. Âûÿâ-
ëåíà íèçêàÿ ÷àñòîòà àëëåëÿ 1475A (0,012) è îòñóòñòâèå àëëåëÿ
1891A â ïîïóëÿöèè Óêðàèíû. Âûâîäû. Ñîçäàíû ìåòîäèêè îáíàðó-
æåíèÿ çàìåí G1475A è G1891A â ãåíå EPHA1 è ïðîâåäåíû ïðåäâà-
ðèòåëüíûå èññëåäîâàíèÿ ðàñïðåäåëåíèÿ ïîëèìîðôèçìîâ G1475A è
G1891A â ïîïóëÿöèè Óêðàèíû.
Êëþ÷åâûå ñëîâà: ãåí EPHA1, ïîëèìîðôèçì, ïîïóëÿöèÿ.
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Received 02.04.13
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