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Histone Abundance Quantification via Flow Cytometry of Htb2-GFP Allows Easy Monitoring of Cell Cycle Perturbations in Living Yeast Cells, Comparable to Standard DNA Staining

J Fungi (Basel). 2023-10; 
Maria V Kulakova, Eslam S M O Ghazy, Fedor Ryabov, Yaroslav M Stanishevskiy, Michael O Agaphonov, Alexander I Alexandrov
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Proteins, Expression, Isolation and Analysis … polymorpha codon-optimized R-GECO encoding sequence was synthesized by GenScript (Piscataway, NJ, USA). This sequence was placed under the control of the OpaMAL1 … Get A Quote

摘要

Assaying changes in the amount of DNA in single cells is a well-established method for studying the effects of various perturbations on the cell cycle. A drawback of this method is the need for a fixation procedure that does not allow for in vivo study nor simultaneous monitoring of additional parameters such as fluorescence of tagged proteins or genetically encoded indicators. In this work, we report on a method of Histone Abundance Quantification (HAQ) of live yeast harboring a GFP-tagged histone, Htb2. We show that it provides data highly congruent with DNA levels, both in and yeasts. The protocol for the DNA content assay was also optimized to be suitable for both and yeasts. Using the HAQ approach, we ... More

关键词

Ogataea, Saccharomyces, cell cycle, flow cytometry, histone, yeast
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