JoVE 비디오를 활용하시려면 도서관을 통한 기관 구독이 필요합니다. 전체 비디오를 보시려면 로그인하거나 무료 트라이얼을 시작하세요.
1INRAe, AgroParisTech, Micalis Institute,Université Paris-Saclay, 2Helmholtz Institute for RNA-based Infection Research (HIRI),Helmholtz-Centre for Infection Research (HZI), 3Medical Faculty,University of Würzburg, 4Centre de Biochimie Structurale, INSERM U1054, CNRS UMR 5048,University of Montpellier
Chapters
- 00:05Introduction
- 00:50Cell Culture and Lysate Preparation
- 04:17Cell-Free Buffer Calibration for Linear DNA
- 05:49Experimental Execution
- 06:33Results: Expression in Cell-Free Protein Synthesis Systems Using Linear DNA Templates
- 07:26Conclusion
Aquí se presenta un protocolo para la preparación y calibración tampón de extractos celulares de cepas knockout de exonucleasa V de Escherichia coli BL21 Rosetta2 (ΔrecBCD y ΔrecB). Este es un enfoque rápido, fácil y directo para la expresión en sistemas de síntesis de proteínas libres de células utilizando plantillas de ADN lineal.
