[논문]Screening and Development of DNA Aptamers Specific to Several Oral Pathogens

Park, Jung-Pyo; Shin, Hye Joo; Park, Suk-Gyun; Oh, Hee-Kyun; Choi, Choong-Ho; Park, Hong-Ju; Kook, Min-Suk; Ohk, Seung-Ho

doi:10.4014/jmb.1407.07019

Screening and Development of DNA Aptamers Specific to Several Oral Pathogens 원문보기

Journal of microbiology and biotechnology, v.25 no.3, 2015년, pp.393 - 398

Park, Jung-Pyo (Department of Oral and Maxillofacial Surgery, Chonnam National University) , Shin, Hye Joo (Department of Oral Microbiology, Chonnam National University) , Park, Suk-Gyun (Department of Oral Microbiology, Chonnam National University) , Oh, Hee-Kyun (Department of Oral and Maxillofacial Surgery, Chonnam National University) , Choi, Choong-Ho (Department of Preventive and Public Health Dentistry, Chonnam National University) , Park, Hong-Ju (Department of Oral and Maxillofacial Surgery, Chonnam National University) , Kook, Min-Suk (Department of Oral and Maxillofacial Surgery, Chonnam National University) , Ohk, Seung-Ho (Department of Oral Microbiology, Chonnam National University)

Abstract ▼ AI-Helper

Aptamers are composed of single-stranded oilgonucleotides that can selectively bind desired molecules. It has been reported that RNA or DNA could act as not only a genetic messenger but also a catalyst in metabolic pathways. RNA aptamers (average sizes 40-50 bp) are smaller than antibodies and have strong binding capacities to target molecules, similar to antigenantibody interactions. Once an aptamer was selected, it can be readily produced in large quantities at low cost. The objectives of this study are to screen and develop aptamers specific to oral pathogens such as Porphyromonas gingivalis, Treponema denticola, and Streptococcus mutans. The bacterial cell pellet was fixed with formaldehyde as a target molecule for the screening of aptamers. The SELEX method was used for the screening of aptamers and a modified western blot analysis was used to verify their specificities. Through SELEX, 40 kinds of aptamers were selected and the specificity of the aptamers to the bacterial cells was confirmed by modified western blot analysis. Through the SELEX method, 40 aptamers that specifically bind to oral pathogens were screened and isolated. The aptamers showed possibility as effective candidates for the detection agents of oral infections.

주제어

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제안 방법

SELEX was carried out by the method of Tuerk and Gold [20] with slight modification. The DNA library was prepared with 44mer and 38mer of primer sequences attached to the 5’ end and 3’ end of randomly synthesized 39 nucleotides.

대상 데이터

Porphyromonas gingivalis ATCC 33277, Treponema denticola ATCC 33521, Streptococcus mutans ATCC 25175, Streptococcus oralis ATCC 10557, and Streptococcus sanguis ATCC 10556 were used in this study. BHI broth containing 0.

성능/효과

Specifically, their ability to bind to a certain protein expressed on the surfaces of the cell or tissues in a certain disease is a useful tool to develop a biosensor or a diagnostic chip for the early detection of the infection or disease. Conclusively, these results presented in this study confirm the possibility of aptamers as a useful tool to detect and diagnosis oral infections. Further studies about the mechanical devices that could make them easily attachable to the human mouth and process the electrical signal to visualize the data should be followed for a complete oral diagnostic system.

후속연구

Conclusively, these results presented in this study confirm the possibility of aptamers as a useful tool to detect and diagnosis oral infections. Further studies about the mechanical devices that could make them easily attachable to the human mouth and process the electrical signal to visualize the data should be followed for a complete oral diagnostic system.
Otherwise, a direct PCR procedure with cell-bound aptamer might amplify the aptamers in an enough amount to clone into the appropriate cloning vector. More high-throughput procedures might be achieved by packing the cell pellet into a small syringe column and letting the aptamer library solution flow through the column.

참고문헌 (21)

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Screening and Development of DNA Aptamers Specific to Several Oral Pathogens 원문보기

Abstract ▼ AI-Helper

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Screening and Development of DNA Aptamers Specific to Several Oral Pathogens 원문보기

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