초록

Ⅳ. 연구개발결과
현재 ‘mixing-spraying’ 기법을 활용한 time-resolved cryo-EM의 구현 단계에 도달하였고, ePEC를 구성하는 효소인 RNAP에 대한 샘플 최적화에 성공하였다.

(출처 : 요약문 4p)

Abstract ▼

Elemental pause is a short (<5 sec) pause that occurs every ~100 nucleotides during transcription. Elemental pause always precedes every gene regulatory process, but its mechanism is not revealed because of the lack of tools capturing the transient intermediates during the elemental pause. Cryo-EM h

Elemental pause is a short (<5 sec) pause that occurs every ~100 nucleotides during transcription. Elemental pause always precedes every gene regulatory process, but its mechanism is not revealed because of the lack of tools capturing the transient intermediates during the elemental pause. Cryo-EM has been a powerful tool for studying the structures of macromolecules, but this conventional method requires tens of seconds for sample preparation. To overcome this limitation, a time-resolved cryo-EM method was developed. The time-resolution of ~ 20 ms has been demonstrated by rapid cooling after mixing and spraying using a microfluidic chip consisting of a passive mixer channel, a reaction channel, and a spray nozzle.
We developed a parylene-based mixing-spraying microfluidic chip, which enables faster stabilization of spray and uniform droplet sizes, significantly reducing sample consumption. Using this microchip, we obtained optimal droplet size distribution and ice thickness on the cryo-EM sample, which allowed clear imaging of the E.coli RNA polymerase and transient transcription elongation complexes. We expect to apply this 'mixing-spraying' method to reveal the mechanism of elemental pause, and further, various biological processes uncovered.

(source : Summary 5p)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 보고서 초록 ... 3
• 요 약 문 ... 4
• SUMMARY ... 5
• CONTENTS ... 6
• 목차 ... 7
• 제 1 장 기본 전사 정지의 정의와 연구 필요성 ... 8
• 제 1 절 기본 전사 정지의 정의 ... 8
• 제 2 절 기본 전사 정지의 연구 현황 및 필요성 ... 8
• 제 2 장 시간 분해 초저온 전자현미경의 국내외 현황 ... 10
• 제 3 장 연구개발수행 내용 및 결과 ... 11
• 제 1 절 연구 목표 및 내용 ... 11
• 제 2 절 연구개발수행 결과 ... 11
• 제 4 장 목표달성도 및 관련분야에의 기여도 ... 14
• 제 5 장 연구개발결과의 활용계획 ... 15
• 제 7 장 참고문헌 ... 16
• 부록. 보충 자료 ... 17
• 끝페이지 ... 21