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Biology
基于蛋白质的继承在酿酒酵母中的高通量筛选
基于蛋白质的继承在酿酒酵母中的高通量筛选
JoVE Journal
Biology
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JoVE Journal Biology
High-throughput Screening for Protein-based Inheritance in S. cerevisiae

基于蛋白质的继承在酿酒酵母中的高通量筛选

Full Text
6,771 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.

此协议说明一种高通量方法来功能屏幕在酿酒酵母中的蛋白质基础继承。

这种高通量酵母表型分析的总体目标是筛选具有传递表达的持久表型记忆的蛋白质,作为基于蛋白质的遗传的代理。这种方法可以帮助回答酵母朊病毒领域中的关键问题,例如,它可以非法遗传多少种蛋白质。该技术的主要优点是您可以以无偏倚的方式筛选许多蛋白质和通路。

通过在 96 摄氏度下在 30 孔板中培养适当的酵母细胞来开始此过程,如文本方案中所述。目视检查培养物以确认培养物已饱和,细胞应在每个孔的底部肉眼可见。使用液体处理机器人制备 384 孔板,每孔含有 45 μL 的适当培养基。

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