A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
- 00:05Title
- 01:46Construction of Strains Containing FFL-GFP Plasmid
- 02:23Preparations Prior to FFL-GFP Induction and Enzymatic Recovery Assay
- 03:48Induction of FFL-GFP
- 06:22Enzymatic FFL-GFP Recovery Assay
- 07:35Fluorescence Microscopy
- 08:55Single Cell Microscopy
- 10:47Results: Hsp104 is Required for Efficient Refolding of Heat-denatured FFL-GFP
- 13:36Conclusion
Denne artikel beskriver anvendelsen af en ildflue luciferase-GFP-fusionsproteinet til at undersøge<em> In vivo</em> Proteinfoldning i<em> Saccharomyces cerevisiae</em>. Brug af denne reagens refoldning af en model varme-denaturerede protein kan monitoreres samtidigt ved fluorescensmikroskopi og en enzymatisk assay til at undersøge roller proteostasis netværkskomponenter i protein kvalitetskontrol.
