Page 32 - tmp

Detection of vitamin D receptor rs2228570 gene variant in patients with systemic
sclerosis

Olgica Maksimović - Regional centre for talents Belgrade II

Introduction

Detection of polymorphism on exon 2 VDR gene was carried
Systemic sclerosis is a chronic disease that is described by the
damage of small blood vessels of the skin (1). Vitamin D has out by PCR-RFLP method. Enzyme FokI recognises GGATG
sequence specifically. Expected product length of digest
an important role in the immune system in which it binds to reaction for the allele F is 266 bp; 193 bp and 73 bp for allele
his receptor VDR (vitamin D receptor) (2,3) therefore taking f and finally 266 bp, 193 bp, 73 bp for heterozygous
part in immunoregulation. The gene variant T>C in SNP combination of alleles Ff for the SNP. Products of restriction
rs2228570 of the VDR gene (FokI polymorphism) imfluences digest reaction were represented by agarose gel
the structure of the VDR protein and it is important in its electrophoresis 2% concentration as well as PCR product not
activity. sliced by restriction enzymes and DNA marker with the
lenght of 100 bp. (Picture 2).
Materials and Methods
Conclusion
Ten patients diagnosed with systemic sclerosis were included
in the research. PCR-RFLP method was used for the detection PCR-RFLP method was applied for the detection of gene
of FokI polymorphism. DNA samples were multiplied by variant T>C within SNP rs2228570 gene for VDR. The given
results show that this method can be used for the analysis of
PCR. Afterwards, the samples were treated with FokI said gene variant. All three variants (FF, Ff and ff) were
restriction enzyme. Depending on the accessibility of the found in the sample of 10 subjects. By examining the whole
restriction sites, it will create DNA fragments with different group of subjects and cross referencing the given data with
lengths. The products of the restriction digest reaction were the control group of healthy subjects we can determine if this
analyzed using agarose gel electrophoresis. gene variant is linked with the occurrence of systemic
sclerosis in the population.
Results and discussion
Literature

1. Clements P.J. Furst D.E Sysytemic sclerosis,
Baltimore, Wiliams and Wilkins 1996.
2. Expression of 1,25-Dihydroxyvitamin D 3 Receptors
in Normal and Psoriatic Skin. Journal of Investigative
Picture 1. Crosscheck of PCR amplification by electrophoresis
on 2% agarose gel. Electrophoretogram of PCR products of 10 Dermatology (1991) 97, 230–239; doi:10.1111/1523-
subjects and its expected length of 266 bp. Position of 1-DNA 1747.ep12480255
marker length of 100bp. Position 2,3,4,5,6,7,8,9,10,11-PCR 3. Mounting Evidence for Vitamin D as an
products Environmental Factor Affecting Autoimmune
Disease Prevalence mpact Factor:2.226
| Ranking:Medicine, Research & Experimental 61 out
of 122 | 5-Year Impact Factor:3.023 | 5-Year
Ranking:Medicine, Research & Experimental 40 out
of 122

Picture 2. Electrophoretogram of PCR-RFLP analysis of SNP
rs2228570 gene VDR. Position of 1-DNA marker with 100 bp.
Position 2, 4, 5, 7, 10 – samples with genotype Ff; 3, 8, 9 – samples
with FF genotype; 6-sample with ff genotype; 11-PCR product not
treated by restriction enzyme.