Selection, Identification, and Binding Mechanism Studies of an ssDNA Aptamer Targeted to Different Stages of E. coli O157:H7

Journal of Agricultural and Food Chemistry
Ying ZouZhouping Wang

Abstract

Enterohemorrhagic Escherichia coli O157:H7 ( E. coli O157:H7) is known as an important food-borne pathogen related to public health. In this study, aptamers which could bind to different stages of E. coli O157:H7 (adjustment phase, log phase, and stationary phase) with high affinity and specificity were obtained by the whole cell-SELEX method through 14 selection rounds including three counter-selection rounds. Altogether, 32 sequences were obtained, and nine families were classified to select the optimal aptamer. To analyze affinity and specificity by flow cytometer, an ssDNA aptamer named Apt-5 was picked out as the optimal aptamer that recognizes different stages of E. coli O157:H7 specifically with the Kd value of 9.04 ± 2.80 nM. In addition, in order to study the binding mechanism, target bacteria were treated by proteinase K and trypsin, indicating that the specific binding site is not protein on the cell membrane. Furthermore, when we treated E. coli O157:H7 with EDTA, the result showed that the binding site might be lipopolysaccharide (LPS) on the outer membrane of E. coli O157:H7.

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Citations

Jul 13, 2019·Critical Reviews in Food Science and Nutrition·Wei WuQingli Yang
Feb 13, 2020·Mini Reviews in Medicinal Chemistry·Weibin Li
Jul 28, 2020·International Journal of Molecular Sciences·Nevina E Trunzo, Ka Lok Hong
Aug 28, 2020·Frontiers in Pharmacology·Morgan L Marshall, Kylie M Wagstaff
Dec 31, 2020·International Journal of Molecular Sciences·Mia StromSarah Shigdar
Jan 29, 2021·Analytical and Bioanalytical Chemistry·Lin ZhuChaoyong Yang
Jan 15, 2021·Comprehensive Reviews in Food Science and Food Safety·Yafang ShenYanbin Li
Mar 9, 2021·Frontiers in Microbiology·Mariam Saad, Sebastien P Faucher
Jun 4, 2021·Veterinary Research·Solène NiederlenderGrégory Karadjian

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