MOLECULAR-GENETIC CHARACTERIZATION OF

ApoE GENE POLYMORPHISM AND

VNTR POLYMORPHISM OF eNOS GENE IN HUMAN

POPULATION OF TUZLA CANTON

1 Department of Biology, Faculty of Natural Sciences and Mathematics, University of Tuzla, B&H

2 Institute of Genetic Engineering and Biotechnology – INGEB, Sarajevo, B&H

INTRODUCTION MATERIALS AND METHODS

Cardiovascular diseases (CVD), as one of the world's leading healthproblems, include a large number of complications that are related togenetic and environmental factors. Understanding the genetics ofCVD origin and development is an important step in preventing andreducing the incidence of CVD. The aim of numerous biomedicalresearches is to identify specific genes and their variants, and theirconnection with CVD.There are a large number of candidate genes that are associated withthe development and emergence of complex CVD, and the moststudied genes were ApoE, ACE, MTHFR, eNOS, 1691 (G>A) FVL,20210 (G>A) PT and polymorphism of 4G/5G PAI-1 gene. Given theimportance of molecular research and diagnostics related to CVD, thedetermination of allelic and genotypic frequencies of ApoE and eNOSgenes in the human population of Tuzla Canton, and their associationwith CVD was analyzed in this paper.

Part of the molecular analysis, isolation of deoxyribonucleic acid(DNA) samples from blood was done in the Laboratory of Geneticsand Microbiology, Department of Biology, Faculty of Science andMathematics, University of Tuzla, while molecular genetic DNA typingwas performed in the Laboratory of Human Genetics at the Instituteof Genetic Engineering and Biotechnology (INGEB), University ofSarajevo.Genotyping of the ApoE gene for all subjects was performed by PCR-RFLP method, according to the protocol (Jovanovic, 2014) which wasfurther optimized, as well by applying a precise variants determinationsystem on the Agilent 2100 Bioanalyzer. Polymerase chain reactionamplified DNA segments of the eNOS gene according to a protocol(Ramirez-Patino et al., 2013) that was further optimized. Moleculargenotyping of the eNOS 4a/b genotype was detected by capillaryelectrophoresis on a genetic analyzer ABI PRISM ®3500 (GeneticAnalyzer 3500, Applied Biosystems).

RESULTS

In the total sample of respondents, the highest relative frequency was recorded for genotype E3/E3 (49.0%), and the

lowest for genotype E2/E4 (1.0%). The E4/E4 genotype was not detected in any group of subjects (Table 1).

The frequency distribution of polymorphisms (alleles and genotypes) of the eNOS gene (4a/4b) was determined for the subjects of the control group as well as for the subjects of the group with cardiovascular diseases (Table 2).

DISCUSSION

The obtained results of ApoE and eNOS gene polymorphism frequency in a sample of

CVD subjects and healthy subjects without known risk factors for CVD development

from Tuzla Canton were compared with data from relevant literature sources in the

studied part of the European and world population (Fatin et al., 2013; Mehmetoğlu et

al., 2010; Djan et al., 2011; Jovanovic, 2014).

CONCLUSION

The results of these studies can be the starting point for further research in molecular

clinical diagnostics, assessment of sensitivity and informativeness of ApoE and eNOS

gene polymorphisms, as a possible adequate potential in clinical and prognostic

implications.

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https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3835499/ [20. januara 2021.]