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JoVE Journal
Biology
Elevado-throughput Screening para herança à base de proteínas no S. cerevisiae
Elevado-throughput Screening para herança à base de proteínas no S. cerevisiae
JoVE Journal
Biology
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JoVE Journal Biology
High-throughput Screening for Protein-based Inheritance in S. cerevisiae

Elevado-throughput Screening para herança à base de proteínas no S. cerevisiae

Full Text
6,768 Views
08:12 min
August 8, 2017

DOI: 10.3791/56069-v

James S. Byers1, Daniel F. Jarosz1,2

1Department of Developmental Biology,Stanford University School of Medicine, 2Department of Chemical and Systems Biology,Stanford University School of Medicine

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol describes a high-throughput methodology to functionally screen for protein-based inheritance in S. cerevisiae. The method aims to identify proteins that can induce a lasting phenotypic memory, providing insights into protein-based inheritance mechanisms.

Key Study Components

Area of Science

  • Neuroscience
  • Cell Biology
  • Genetics

Background

  • Protein-based inheritance is a key area of research in yeast.
  • This study focuses on the role of proteins in phenotypic memory.
  • Understanding these mechanisms can shed light on prion biology.
  • The methodology allows for unbiased screening of multiple proteins.

Purpose of Study

  • To screen for proteins that can elicit a lasting phenotypic memory.
  • To investigate the potential for protein-based inheritance in yeast.
  • To provide a high-throughput approach for studying protein functions.

Methods Used

  • Yeast cells are grown in a 96 well plate at 30 degrees Celsius.
  • Cultures are visually inspected for saturation.
  • A liquid handling robot prepares 384 well plates with the appropriate medium.
  • Screening is conducted in an unbiased manner across various proteins.

Main Results

  • The method allows for the identification of proteins that induce phenotypic memory.
  • It provides insights into the number of proteins capable of such inheritance.
  • The high-throughput nature of the study enhances the screening process.
  • Results contribute to understanding the yeast prion field.

Conclusions

  • This protocol offers a novel approach to studying protein-based inheritance.
  • The findings may have implications for broader biological research.
  • Future studies can build on this methodology to explore additional proteins.

Frequently Asked Questions

What is the main goal of this study?
The main goal is to functionally screen for proteins that can induce lasting phenotypic memory in yeast.
How does this method improve upon previous techniques?
This method allows for high-throughput, unbiased screening of multiple proteins simultaneously.
What organism is used in this research?
The research is conducted using the yeast species S. cerevisiae.
What temperature is used for growing yeast cultures?
Yeast cultures are grown at 30 degrees Celsius.
What is the significance of studying protein-based inheritance?
Studying protein-based inheritance can provide insights into mechanisms of memory and inheritance in biological systems.
What type of plates are used in the screening process?
The screening process utilizes 96 well and 384 well plates for culturing and testing.

Este protocolo descreve uma metodologia de alta produtividade para a tela funcionalmente para herança à base de proteínas no S. cerevisiae.

O objetivo geral desta fenotipagem de levedura de alto rendimento é rastrear proteínas que ilícitas uma memória fenotípica duradoura de expressão passada, como um proxy para a herança baseada em proteínas. Este método pode ajudar a responder a perguntas-chave no campo do príon de levedura, como, por exemplo, quantas proteínas podem herança ilícita desse tipo. A principal vantagem dessa técnica é que você pode rastrear muitas proteínas e vias de maneira imparcial.

Inicie este procedimento cultivando as células de levedura apropriadas em uma placa de 96 poços a 30 graus Celsius, conforme descrito no protocolo de texto. Inspecione visualmente as culturas para confirmar se as culturas estão saturadas, as células devem ser visíveis a olho nu, no fundo de cada poço. Use um robô de manuseio de líquidos para preparar placas de 384 poços, contendo 45 microlitros do meio apropriado por poço.

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