Cargando…

Protocol for Titrating Gene Expression Levels in Budding Yeast

The biological phenotype is affected by the level of gene expression. Here, we provide a step-by-step protocol for precisely titrating and quantitatively observing the target gene expression level in budding yeast by manipulating its copy number in the genome. Using this method, we construct various...

Descripción completa

Detalles Bibliográficos
Autores principales: Qu, Yimiao, Jiang, Jun, Yang, Xiaojing, Tang, Chao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580245/
https://www.ncbi.nlm.nih.gov/pubmed/33111115
http://dx.doi.org/10.1016/j.xpro.2020.100082
Descripción
Sumario:The biological phenotype is affected by the level of gene expression. Here, we provide a step-by-step protocol for precisely titrating and quantitatively observing the target gene expression level in budding yeast by manipulating its copy number in the genome. Using this method, we construct various strains with different gene copy numbers of the cell cycle inhibitor Whi5. This protocol enables stable and inherent control of gene expression at the expected level with fluorescent intensity as the quantitative readout. For complete details on the use and execution of this protocol, please refer to Qu et al. (2019).