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논문 기본 정보

자료유형
학술저널
저자정보
Shim Jiyoung (Department of Chemistry Western Kentucky University Bowling Green KY 42101 USA) Williams Langley (Department of Chemistry Western Kentucky University Bowling Green KY 42101 USA) Dohyun Kim (Department of Mechanical Engineering Myongji University Yongin 449-728 Republic of Korea) Ko Kisung (Department of Medicine College of Medicine Chung-Ang University Seoul 06974 Republic of Korea) Kim Moon-Soo (Department of Chemistry Western Kentucky University Bowling Green KY 42101 USA)
저널정보
한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제31권 제9호
발행연도
2021.9
수록면
1,323 - 1,329 (7page)
DOI
10.4014/jmb.2106.06057

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Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a doublestranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.

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