Development and Characterization of Aptamer for rRNA Methyltransferase Enzyme in Aminoglycoside Resistant Bacteria
Project no. 23068 · Status: Kapanmış (Yayın bekleniyor) · 2021
Summary
<p>The World Health Organization has listed aminoglycoside antibiotics as critically important antimicrobials for treating bacterial infections, including tuberculosis, endocardial and meningeal infections, and plague. Due to their ability to be transferred via plasmids, mechanisms of resistance against these antibiotics can spread rapidly among bacteria. Therefore, monitoring aminoglycoside resistance mechanisms has been identified as a key focus of this study. Accordingly, the aim of the study is to develop aptamers as molecular recognition element for detecting aminoglycoside resistance. The 16S rRNA methyltransferase enzyme plays a significant role in aminoglycoside resistance by methylating the ribosome, thereby preventing the inhibitory effect of aminoglycosides on translation. Detecting and monitoring the presence of this enzyme allows for the identification and tracking of resistance. Therefore, the development of aptamers for detecting or monitoring aminoglycoside resistance is aimed at addressing these reasons.</p><p>Within the thesis study, the enzyme 16S rRNA methyltransferase, which causes resistance, was cloned into <em>E. coli</em> T7 to obtain the active enzyme. Using the SELEX method, the aptamer library was screened through 14 rounds, and groups of aptamers thought to be suitable for the enzyme were sent for sequencing. After the bioinformatic analysis of the sequences, two aptamers were identified, and binding analyses were conducted. The EMSA analysis demonstrated that the A1 and A2 aptamers specifically bind to the armA protein. The binding resulted in a Kd value of 951 ppm for the A1 aptamer and 1014 ppm for the A2 aptamer.</p>