Department of Biochemistry and Molecular Pharmacology, New York University Langone School of Medicine, New York, NY 10016, USA.
Institute for Systems Genetics, New York University Langone School of Medicine, New York, NY 10016, USA.
Science. 2017 Mar 10;355(6329). doi: 10.1126/science.aaf4831.
We describe design, rapid assembly, and characterization of synthetic yeast Sc2.0 chromosome VI (synVI). A mitochondrial defect in the synVI strain mapped to synonymous coding changes within (), encoding an essential proteasome subunit; Sc2.0 coding changes reduced Pre4 protein accumulation by half. Completing Sc2.0 specifies consolidation of 16 synthetic chromosomes into a single strain. We investigated phenotypic, transcriptional, and proteomewide consequences of Sc2.0 chromosome consolidation in poly-synthetic strains. Another "bug" was discovered through proteomic analysis, associated with alteration of the transcription start due to transfer RNA deletion and loxPsym site insertion. Despite extensive genetic alterations across 6% of the genome, no major global changes were detected in the poly-synthetic strain "omics" analyses. This work sets the stage for completion of a designer, synthetic eukaryotic genome.
我们描述了合成酵母 Sc2.0 染色体 VI（synVI）的设计、快速组装和特性分析。synVI 菌株的线粒体缺陷映射到 （）内的同义编码变化，该基因编码一个必需的蛋白酶体亚基；Sc2.0 的编码变化将 Pre4 蛋白的积累减少了一半。完成 Sc2.0 后，将 16 条合成染色体整合到一个单一的菌株中。我们研究了多合成菌株中 Sc2.0 染色体整合的表型、转录和全蛋白质组学后果。通过蛋白质组学分析发现了另一个“错误”，它与转移 RNA 缺失和 loxPsym 位点插入导致的 转录起始改变有关。尽管基因组的 6%发生了广泛的遗传改变，但在多合成菌株的“组学”分析中没有检测到主要的全局变化。这项工作为完成一个设计的、合成的真核基因组奠定了基础。
