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DETEKCIJA GENSKE VARIJANTE RS2228570 RECEPTORA ZA VITAMIN D KOD
PACIJENATA OBOLELIH OD SISTEMSKE SKLEROZE

DETECTION OF VITAMIN D RECEPTOR RS2228570 GENE VARIANT IN PATIENTS
WITH SYSTEMIC SCLEROSIS

Autor: Olgica Maksimović Učenica I razreda Treće beogradske gimnazije

Regionalni Centar za talente Beograd II
Mentori: dr Vesna Spasovski, nauĉni saradnik Institut za molekularnu genetiku i genetiĉko

inţenjerstvo ( IMGGI ) Maja Gajić, student Biološkog fakulteta

REZIME

Sistemska skleroza je hronična autoimuna bolest koju karakteriše oštedenje malih krvnih sudova kože. Važna uloga
vitamina D je u imunskom sistemu gde svoju imunoregulatornu ulogu ostvaruje putem vezivanja za svoj receptor
VDR (vitamin D receptor). Genska varijanta T>C u oviru SNPrs2228570 u VDR genu (FokI polimorfizam) utiče na
strukturu VDR proteina i značajna je za njegovu aktivnost. U ispitivanje je uključeno 10 bolesnika obolelih od
sistemske skleroze. Za detekciju FokI polimorfizma korišdena je metoda PCR-RFLP. Uzorci DNK umnoženi su PCR
metodom, a zatim tretirani restrikcionim enzimom FokI. Enzim FokI specifično prepoznaje GGATG sekvencu.
Očekivana dužina produkta digestije za alel F je 266bp; 193bp i 73bp za alel f, odnosno 266bp, 193bp i 73bp za
heterozigotnu kombinaciju Ff za ovaj SNP. Produkti digestije vizuelizovani su elektroforezom na agaroznom gelu.
Dobijeni rezultati potvrđuju da se ova metoda može koristiti za analizu ove genske varijante. Ispitivanje na uzorku
od 10 pacijenata utvrđeno je prisustvo sve tri varijante: FF, Ff, i ff. Ispitivanjem cele grupe ispitanika i poređenjem
uzoraka pacijenata sa kontrolnom grupom zdravih ispitanika utvrdide se da li je ova genska varijanta povezana sa
nastankom sistemske skleroze u našoj populaciji.

Ključne reči: sistemska skleroza, vitamin D, FokI polimorfizam, VDR gen, PCR-RFLP

SUMMERY

Systemic sclerosis is a chronic disease that is described by the damage of small blood vessels of the skin. Vitamin D
has an important role in the immune system in which it binds to his receptor VDR (vitamin D receptor) therefore
taking part in immunoregulation. The gene variant T>C in SNP rs2228570 of the VDR gene (FokI polymorphism)
imfluences the structure of the VDR protein and it is important in its activity. Ten patients diagnosed with systemic
sclerosis were included in the research. PCR-RFLP method was used for the detection of FokI polymorphism. DNA
samples were multiplied by PCR. Afterwards, the samples were treated with FokI restriction enzyme. Enzyme FokI
recognises GGATG sequence specifically. Expected product length of digest reaction for the allele F is 266 bp; 193
bp and 73 bp for allele f and finally 266 bp, 193 bp, 73 bp for heterozygous combination of alleles Ff for the SNP.
Products of restriction digest reaction were represented by agarose gel electrophoresis. The given results show
that this method can be used for the analysis of said gene variant. All three variants (FF, Ff and ff) were found in the
sample of 10 subjects. By examining the whole group of subjects and cross referencing the given data with the
control group of healthy subjects we can determine if this gene variant is linked with the occurrence of systemic
sclerosis in the population.

Key words: Systemic sclerosis, vitamin D, FokI polymorphism, VDR gene, PCR-RFLP