莱茵衣藻
转化(遗传学)
可选择标记
生物
酿酒酵母
线粒体DNA
遗传学
同源重组
线粒体
基因组
基因
计算生物学
突变体
作者
Nathalie Bonnefoy,Claire Remacle
出处
期刊:Methods in molecular biology
日期:2023-01-01
卷期号:: 345-364
被引量:2
标识
DOI:10.1007/978-1-0716-2922-2_24
摘要
Chlamydomonas reinhardtii and Saccharomyces cerevisiae are currently the two micro-organisms in which genetic transformation of mitochondria is routinely performed. The generation of a large variety of defined alterations as well as the insertion of ectopic genes in the mitochondrial genome (mtDNA) are possible, especially in yeast. Biolistic transformation of mitochondria is achieved through the bombardment of microprojectiles coated with DNA, which can be incorporated into mtDNA thanks to the highly efficient homologous recombination machinery present in S. cerevisiae and C. reinhardtii organelles. Despite a low frequency of transformation, the isolation of transformants in yeast is relatively quick and easy, since several natural or artificial selectable markers are available, while the selection in C. reinhardtii remains long and awaits new markers. Here, we describe the materials and techniques used to perform biolistic transformation, in order to mutagenize endogenous mitochondrial genes or insert novel markers into mtDNA. Although alternative strategies to edit mtDNA are being set up, so far, insertion of ectopic genes relies on the biolistic transformation techniques.
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