Recombinant Production of E. coli NAD+-dependent DNA ligase as a Target for Antibacterial Drug Discovery

Özlem Kaplan; Rizvan İmamoğlu; İsa Gökçe

doi:10.18016/ksutarimdoga.vi.884279

TR EN

Antibakteriyel İlaç Keşfi İçin Bir Hedef Olarak E. Coli NAD+-Bağımlı DNA Ligazın Rekombinant Üretimi

Öz

Bakteriyel enfeksiyonlarda ilaç direnci frekansındaki artış, yeni mekanizmaları hedefleyen yeni antibakteriyel ajanların araştırılmasına yol açmıştır. NAD+-bağımlı DNA ligazın fonksiyonlarının bir çoğu, onu antibakteriyel ilaç keşfi için dikkate değer bir hedef haline getirmiştir. Escherichia coli (E. coli) NAD+ bağımlı DNA ligazı, ATP bağımlı insan DNA ligazı ile karşılaştrıldığında, benzersiz substrat özgüllüğü nedeniyle potansiyel bir hedef olarak görülmektedir. Bu çalışmada antibakteriyel ilaç keşiflerinde sıklıkla kullanılan E. coli NAD+ bağımlı DNA ligaz enziminin, yüksek miktarda ve saflıkta üretilmesi amaçlamıştır. E. coli DNA ligaz gen dizisi, pTOLT vektör sistemine klonlanmıştır. E. coli DNA ligaz enzimi, E. coli BL21 (DE3) pLysE hücrelerinde üretildikten sonra saflaştırılmıştır. Bu çalışmada üretilen DNA ligaz enziminin DNA fragmanlarının ligasyonunu sağlayabildiği, aktivite testi ile açıkça gösterilmiştir. Sonuç olarak, bu enzim üzerinde aday inhibitörlerin etkisinin basitçe incelenebileceği ortaya koyulmuştur.

Anahtar Kelimeler

Recombinant Production of E. coli NAD+-dependent DNA ligase as a Target for Antibacterial Drug Discovery

Abstract

The increase in the frequency of drug resistance in bacterial infections has led to the research of new antibacterial agents targeting new mechanisms. Many of the functions of NAD+-dependent DNA ligase have made it a remarkable target for antibacterial drug discovery. Escherichia coli (E. coli) NAD+-dependent DNA ligase is presented as a potential target due to its unique substrate specificity compared to the ATP-dependent human DNA ligase. In this study, it was aimed to produce and purify the E. coli NAD + dependent DNA ligase enzyme, which is frequently used in antibacterial drug discovery. The E. coli DNA ligase gene sequence was cloned into pTOLT vector system. E. coli DNA ligase enzyme was purified after the production in E. coli BL21 (DE3) pLysE cells. It was clearly demonstrated by the activity test that the DNA ligase enzyme produced in this study can ligate the DNA fragments. As a result, it was revealed that the effect of candidate inhibitors can be studied simply on the enzyme.

Keywords

Supporting Institution

Turkish Scientific and Technical Research Council (TUBITAK)

Project Number

2209-A

Thanks

This study was supported by the Turkish Scientific and Technical Research Council (TUBITAK) (TUBITAK-2209A)

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Details

Primary Language

English

Subjects

-

Journal Section

Research Article

Authors

Özlem Kaplan ^*
0000-0002-3052-4556
Türkiye

Rizvan İmamoğlu
0000-0002-6306-4760
Türkiye

İsa Gökçe
0000-0002-5023-9947
Türkiye

Publication Date

February 28, 2022

Submission Date

February 21, 2021

Acceptance Date

April 20, 2021

Published in Issue

Year 2022 Volume: 25 Number: 1

DOI

https://doi.org/10.18016/ksutarimdoga.vi.884279

IZ

https://izlik.org/JA32HD45MG

Cite

RIS / Bibtex

APA

Kaplan, Ö., İmamoğlu, R., & Gökçe, İ. (2022). Recombinant Production of E. coli NAD+-dependent DNA ligase as a Target for Antibacterial Drug Discovery. Kahramanmaraş Sütçü İmam Üniversitesi Tarım Ve Doğa Dergisi, 25(1), 19-24. https://doi.org/10.18016/ksutarimdoga.vi.884279

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