Wu Yi, Li Bing-Zhi, Zhao Meng, Mitchell Leslie A, Xie Ze-Xiong, Lin Qiu-Hui, Wang Xia, Xiao Wen-Hai, Wang Ying, Zhou Xiao, Liu Hong, Li Xia, Ding Ming-Zhu, Liu Duo, Zhang Lu, Liu Bao-Li, Wu Xiao-Le, Li Fei-Fei, Dong Xiu-Tao, Jia Bin, Zhang Wen-Zheng, Jiang Guo-Zhen, Liu Yue, Bai Xue, Song Tian-Qing, Chen Yan, Zhou Si-Jie, Zhu Rui-Ying, Gao Feng, Kuang Zheng, Wang Xuya, Shen Michael, Yang Kun, Stracquadanio Giovanni, Richardson Sarah M, Lin Yicong, Wang Lihui, Walker Roy, Luo Yisha, Ma Ping-Sheng, Yang Huanming, Cai Yizhi, Dai Junbiao, Bader Joel S, Boeke Jef D, Yuan Ying-Jin
Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, PR China.
SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin University, Tianjin, 300072, PR China.
Science. 2017 Mar 10;355(6329). doi: 10.1126/science.aaf4706.
Debugging a genome sequence is imperative for successfully building a synthetic genome. As part of the effort to build a designer eukaryotic genome, yeast synthetic chromosome X (synX), designed as 707,459 base pairs, was synthesized chemically. SynX exhibited good fitness under a wide variety of conditions. A highly efficient mapping strategy called pooled PCRTag mapping (PoPM), which can be generalized to any watermarked synthetic chromosome, was developed to identify genetic alterations that affect cell fitness ("bugs"). A series of bugs were corrected that included a large region bearing complex amplifications, a growth defect mapping to a recoded sequence in , and a loxPsym site affecting promoter function of PoPM is a powerful tool for synthetic yeast genome debugging and an efficient strategy for phenotype-genotype mapping.
调试基因组序列对于成功构建合成基因组至关重要。作为构建设计真核基因组努力的一部分，设计为707,459个碱基对的酵母合成染色体X（synX）通过化学方法合成。SynX在多种条件下表现出良好的适应性。开发了一种名为混合PCR标签映射（PoPM）的高效映射策略，该策略可推广到任何有标记的合成染色体，以识别影响细胞适应性的遗传改变（“错误”）。纠正了一系列错误，包括一个带有复杂扩增的大区域、一个映射到编码序列中的生长缺陷以及一个影响启动子功能的loxPsym位点。PoPM是合成酵母基因组调试的强大工具和表型-基因型映射的有效策略。
