È necessario avere un abbonamento a JoVE per visualizzare questo. Accedi o inizia la tua prova gratuita.
Capitoli
- 00:00Titolo
- 00:31Introduction
- 01:05Preparing the pPICZaA Construct
- 02:18Preparation of Electrocompetent Yeast Cells
- 04:04Transformation by Electroporation
- 06:06Protein Expression in Pichia pastoris
- 08:42Results of Recombinant Protein Expression
- 09:02Conclusion
Protokollet beskriver protein uttryck med methylotrophic jäst<em> Pichia pastoris</em>. Beredningen av electrocompetent jästceller, omvandling av vektorn med genen av intresse i<em> P. pastoris</em> Och jäst DNA-rening utförs också. Western blot analys och proteinrening bygga de sista stegen i denna protein expression protokoll.
Tags
Recombinant ProteinsPichia PastorisProtein ExpressionMethylotrophic YeastExpression SystemPost-translational ModificationsProtein FoldingProteolytic ProcessingDisulfide Bond FormationGlycosylationMethanol MetabolismAlcohol Oxidase PromoterMethanol InductionSecretion Of Recombinant ProteinSignal SequenceHeterologous ProteinProtein PurificationVector
