用于测量酵母染色体丢失的染色体传递保真度检测法。

The Chromosome Transmission Fidelity Assay for Measuring Chromosome Loss in Yeast.

作者信息

Duffy Supipi, Hieter Philip

机构信息

Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada.

出版信息

Methods Mol Biol. 2018;1672:11-19. doi: 10.1007/978-1-4939-7306-4_2.

DOI:10.1007/978-1-4939-7306-4_2

PMID:29043613

Abstract

The budding yeast Saccharomyces cerevisiae has served as an excellent model system for studying highly conserved biological pathways including pathways involved in genome transmission and maintenance. The Chromosome Transmission Fidelity (CTF) colony color assay was developed to assess chromosome instability (CIN) in yeast, by monitoring the loss or gain during cell division of an artificial chromosome fragment carrying a visual marker. The CTF assay monitors changes in chromosome number, allowing the detection of mutants that exhibit increased rates of chromosome nondisjunction or chromosome loss. In this article, we describe the SUP11-marker-based CTF assay system, and the methodologies for both qualitative analysis of mutants affecting chromosome transmission, and quantitative analysis for determining the types and rates of errors in chromosome transmission using half-sector analysis.

摘要

出芽酵母酿酒酵母一直是研究高度保守生物途径（包括参与基因组传递和维持的途径）的优秀模型系统。染色体传递保真度（CTF）菌落颜色测定法是通过监测携带视觉标记的人工染色体片段在细胞分裂过程中的丢失或增加来评估酵母中的染色体不稳定性（CIN）。CTF测定法监测染色体数目的变化，从而能够检测出表现出染色体不分离或染色体丢失率增加的突变体。在本文中，我们描述了基于SUP11标记的CTF测定系统，以及用于对影响染色体传递的突变体进行定性分析和使用半扇形分析确定染色体传递错误类型和速率的定量分析方法。