Melamin, siyanürik asit ve ürik asidin DNA ile etkileşimlerinin incelenmesi
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Dosyalar
Tarih
2020
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Kırıkkale Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
DNA molekülü, çift sarmallı yapıya sahip olması nedeni ile ilaçlar, metal kompleksleri ve birçok molekül için potansiyel bağlanma bölgeleri taşımaktadır. Bağlanmak için DNA'yı hedef olarak seçen söz konusu moleküllerin gerek zararsız hale getirilebilmeleri gerekse olası ilaç etkinliklerinin artırılması için moleküller arası etkileşim şekillerinin aydınlatılması oldukça önemlidir. Azotlu organik bir bileşik olan melamin, metabolizmada ürik asit ve siyanürik asit gibi çeşitli metabolitlere dönüşebilmektedir. Melamin ve metabolitlerinin gastrointestinal sistemde emildiği ve hayvanlarda nefrolitiazis, kronik böbrek iltihabı ve mesane karsinomu dahil olmak üzere çeşitli zararlı etkilere sebep olabildiği ileri sürülmüştür. Bu çalışmada, yüksek azot içeriği sebebiyle süt, süt tozu, bebek mamaları, pet ve çiftlik hayvan yemleri gibi çeşitli gıdalara hile amaçlı katılan melamin ve metabolize olan melaminden oluşan siyanürik asit ve ürik asidin ökaryotik (sığır timus dokusu) ve prokaryotik (Staphylococcus aureus bakteri kültürü) kaynaklardan izole edilen genomik DNA ile olan etkileşimlerinin incelenmesi amaçlandı. Söz konusu etkileşimlerin değerlendirilmesinde UV-absorbans spektrofotometri, floresans spektrofotometri ve agaroz jel elektroforezi tekniklerinden yararlanıldı. Sabit DNA konsantrasyonu ile inkübe edilen, beş farklı konsantrasyondaki melamin, siyanürik asit ve ürik asit çözeltilerinin gerçekleştirilen analizleri neticesinde söz konusu bileşiklerin DNA molekülleri ile etkileşim içerisinde olduğu belirlendi. UV-absorbans ve floresans spektrofotometri analizleri ile elde edilen grafiklerde, artan madde konsantrasyonuna karşın dalga piklerinde de artış gözlendi. Agaroz jel elektoroforez analizleri ile elde edilen jel görüntülerinin piksel ölçümü yapıldığında etkileri incelenen metabolitlerin DNA bant hacminde azalmaya sebep olduğu ve bu durumun, melamin ile hazırlanan kombinasyonlarda daha da belirgin hale geldiği görüldü. Elde edilen bulgular değerlendirildiğinde, melamin, siyanürik asit ve ürik asidin incelenen ökaryotik ve prokaryotik genomik DNA materyallerine groove bağlanma ile bağlandığı sonucuna varıldı. Anahtar Kelimeler: DNA, melamin, siyanürik asit, ürik asit, UV-absorbans, floresans spektrofotometri, agaroz jel elektroforezi
Since the DNA molecule has a double helix structure, it bears potential binding sites for drugs, metal complexes and many molecules. It is very important to enlighten the interactions between molecules choosing DNA as the target to connect in order to make these molecules harmless as well as to increase the drug efficacy. Melamine, an organic compound with nitrogen, can be converted into various metabolites such as uric acid and cyanuric acid in metabolism. It has been suggested that melamine and its metabolites are absorbed in the gastrointestinal tract and can cause various harmful effects in animals, including nephrolithiasis, chronic kidney inflammation and bladder carcinoma. In this study, it was aimed to investigate the interactions of genomic DNA isolated from eukaryotic (bovine thymus tissue) and prokaryotic (bacterial culture of Staphylococcus aureus) sources with melamine which is added to various foods such as milk, milk powder, baby food, pet and livestock feed due to its high nitrogen content for cheating purposes and cyanuric and uric acids which occur from metabolised melamine. UV-absorbance spectrophotometry, fluorescence spectrophotometry and agarose gel electrophoresis techniques were used to evaluate these interactions. As a result of the analysis of the five different concentrations of melamine, cyanuric acid and uric acid solutions incubated with fixed DNA concentration, it was determined that the investigated compounds interact with DNA molecules. In the graphics obtained by UV-absorbance and fluorescence spectrophotometry analysis, an increase in wave peaks was also observed despite the increasing substance concentration. When pixel measurement of gel images obtained by agarose gel electrophoresis analysis was performed, it was observed that the metabolites of which the effects were examined caused a decrease in the DNA band volume and this situation became more evident in combinations prepared with melamine. When the findings obtained are evaluated, it was concluded that melamine, cyanuric acid and uric acid were bonded to the eukaryotic and prokaryotic genomic DNA materials examined through groove binding. Keywords: DNA, melamine, cyanuric acid, uric acid, UV-absorbance, fluorescence spectrophotometry, agarose gel electrophoresis
Since the DNA molecule has a double helix structure, it bears potential binding sites for drugs, metal complexes and many molecules. It is very important to enlighten the interactions between molecules choosing DNA as the target to connect in order to make these molecules harmless as well as to increase the drug efficacy. Melamine, an organic compound with nitrogen, can be converted into various metabolites such as uric acid and cyanuric acid in metabolism. It has been suggested that melamine and its metabolites are absorbed in the gastrointestinal tract and can cause various harmful effects in animals, including nephrolithiasis, chronic kidney inflammation and bladder carcinoma. In this study, it was aimed to investigate the interactions of genomic DNA isolated from eukaryotic (bovine thymus tissue) and prokaryotic (bacterial culture of Staphylococcus aureus) sources with melamine which is added to various foods such as milk, milk powder, baby food, pet and livestock feed due to its high nitrogen content for cheating purposes and cyanuric and uric acids which occur from metabolised melamine. UV-absorbance spectrophotometry, fluorescence spectrophotometry and agarose gel electrophoresis techniques were used to evaluate these interactions. As a result of the analysis of the five different concentrations of melamine, cyanuric acid and uric acid solutions incubated with fixed DNA concentration, it was determined that the investigated compounds interact with DNA molecules. In the graphics obtained by UV-absorbance and fluorescence spectrophotometry analysis, an increase in wave peaks was also observed despite the increasing substance concentration. When pixel measurement of gel images obtained by agarose gel electrophoresis analysis was performed, it was observed that the metabolites of which the effects were examined caused a decrease in the DNA band volume and this situation became more evident in combinations prepared with melamine. When the findings obtained are evaluated, it was concluded that melamine, cyanuric acid and uric acid were bonded to the eukaryotic and prokaryotic genomic DNA materials examined through groove binding. Keywords: DNA, melamine, cyanuric acid, uric acid, UV-absorbance, fluorescence spectrophotometry, agarose gel electrophoresis
Açıklama
Anahtar Kelimeler
Biyokimya, Biochemistry ; Veteriner Hekimliği, Veterinary Medicine
