Probing Transcriptional Dynamics in Living Cells by Single-Molecule Fluorescence Imaging
소속 :
연사 : 이남기 교수 (포항공대 물리학과 & 시스템생명공학부)
일시 : 2009-04-30 17:00 ~
장소 : 500동 목암홀
일시: 2009년 4월 30일 오후 5:00
장소: 500동 목암홀
- Abstract -
Recent advances in single-molecule fluorescence microscope techniques have allowed single-molecule sensitivity to probe various protein-DNA interactions, their structural changes, and fundamental cellular processes in a living cell. In this talk, I will introduce a novel single-molecule fluorescence microscope technique for in vivo single protein detection. To study transcription, the process of DNA-directed RNA biosynthesis in living cells, I used localization-enhancement fluorescence detection technique. I used a fluorescently tagged T7 RNA polymerase in bacteria containing a T7 promoter integrated into the chromosome. Since there are no other endogenous T7 promoters in the E. coli genome, localized T7 polymerase molecules correspond to transcription at the engineered promoter. In this way, I will demonstrate that it is possible to detect single RNA polymerase in living cells and directly measured promoter searching time, transcription initiation and elongation rates of T7 RNA polymerase. This work paves the way to observe individual protein machinery to reveal fundamental cellular processes in living cells.
장소: 500동 목암홀
- Abstract -
Recent advances in single-molecule fluorescence microscope techniques have allowed single-molecule sensitivity to probe various protein-DNA interactions, their structural changes, and fundamental cellular processes in a living cell. In this talk, I will introduce a novel single-molecule fluorescence microscope technique for in vivo single protein detection. To study transcription, the process of DNA-directed RNA biosynthesis in living cells, I used localization-enhancement fluorescence detection technique. I used a fluorescently tagged T7 RNA polymerase in bacteria containing a T7 promoter integrated into the chromosome. Since there are no other endogenous T7 promoters in the E. coli genome, localized T7 polymerase molecules correspond to transcription at the engineered promoter. In this way, I will demonstrate that it is possible to detect single RNA polymerase in living cells and directly measured promoter searching time, transcription initiation and elongation rates of T7 RNA polymerase. This work paves the way to observe individual protein machinery to reveal fundamental cellular processes in living cells.
