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  • 09월 04일 17시 이후 : 초록수정 불가능, 일정확인 및 검색만 가능

제114회 대한화학회 학술발표회, 총회 및 기기전시회 안내 A graphene oxide based platform for detection of mutant fusion DNA using quencher-free fluorescent DNA probe and polymerase chain reaction

2014년 9월 3일 15시 47분 21초
BIO.P-651 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
10월 15일 (수요일) 16:00~19:00
저자 및
노경민, 김동은*
건국대학교 생명공학과, Korea
Graphene oxide (GO) has been applied in diverse biomedical areas as a versatile material for sensors. GO preferentially binds to single-stranded DNA (ssDNA) over double-stranded DNA (dsDNA) and quenches fluorescence when the fluorophore stays close to GO surface. In this study, we developed a simple and efficient method for detection of mutant fusion gene using fluorescent probe DNA and polymerase chain reaction. Bcr-Abl fusion gene that causes Chronic Myeloid Leukemia (CML) was used as a model system for detection of mutant gene. In our designed system for mutant gene detection, GO quenches the fluorescent probe DNA and Taq polymerase degrades the probe DNA due to its 5′-end nuclease activity. When the Bcr-Abl mutant gene is present, PCR amplification of Bcr-Abl results in degradation of the fluorescent probe DNA annealed to the Bcr-Abl amplicon. In the absence of Bcr-Abl, the fluorescent ssDNA probe remains intact due to the absence of amplification. Addition of GO to each reaction solution gave rise to different fluorescence signal; enhanced fluorescence with degraded probe DNA in Bcr-Abl (+), and quenched fluorescence with ssDNA adsorbed onto GO in Bcr-Abl (-). We observed fluorescence signal difference between Bcr-Abl positive and negative cells, and the fluorescence signal was quantitatively correlated with composition of these cells. Thus, our detection system can be applicable as quantitative assay for primary diagnosis as well as monitoring therapeutic responses by measuring the expression level of Bcr-Abl fusion gene in leukemia patients.