Higuchi-Sanabria Ryo, Garcia Enrique J, Tomoiaga Delia, Munteanu Emilia L, Feinstein Paul, Pon Liza A
Department of Pathology and Cell Biology, Columbia University, New York, NY, United States of America.
Department of Biological Sciences, Hunter College and The Graduate Center Biochemistry, Biology and Biopsychology and Behavioral Neuroscience Programs, CUNY, New York, NY 10065, United States of America.
PLoS One. 2016 Jan 4;11(1):e0146120. doi: 10.1371/journal.pone.0146120. eCollection 2016.
Saccharomyces cerevisiae are widely used for imaging fluorescently tagged protein fusions. Fluorescent proteins can easily be inserted into yeast genes at their chromosomal locus, by homologous recombination, for expression of tagged proteins at endogenous levels. This is especially useful for incorporation of multiple fluorescent protein fusions into a single strain, which can be challenging in organisms where genetic manipulation is more complex. However, the availability of optimal fluorescent protein combinations for 3-color imaging is limited. Here, we have characterized a combination of fluorescent proteins, mTFP1/mCitrine/mCherry for multicolor live cell imaging in S. cerevisiae. This combination can be used with conventional blue dyes, such as DAPI, for potential four-color live cell imaging.
酿酒酵母被广泛用于对荧光标记的蛋白质融合体进行成像。荧光蛋白可通过同源重组轻松插入酵母基因的染色体位点，以便在内源水平表达标记蛋白。这对于将多个荧光蛋白融合体整合到单个菌株中尤为有用，而在基因操作更为复杂的生物体中，这可能具有挑战性。然而，用于三色成像的最佳荧光蛋白组合的可用性有限。在这里，我们已经鉴定出一种荧光蛋白组合，即mTFP1/mCitrine/mCherry，用于酿酒酵母中的多色活细胞成像。这种组合可与传统蓝色染料（如DAPI）一起使用，用于潜在的四色活细胞成像。
