Bug mapping and fitness testing of chemically synthesized chromosome X

Yi Wu, Bing Zhi Li, Meng Zhao, Leslie A. Mitchell, Ze Xiong Xie, Qiu Hui Lin, Xia Wang, Wen Hai Xiao, Ying Wang, Xiao Zhou, Hong Liu, Xia Li, Ming Zhu Ding, Duo Liu, Lu Zhang, Bao Li Liu, Xiao Le Wu, Fei Fei Li, Xiu Tao Dong, Bin JiaWen Zheng Zhang, Guo Zhen Jiang, Yue Liu, Xue Bai, Tian Qing Song, Yan Chen, Si Jie Zhou, Rui Ying Zhu, Feng Gao, Zheng Kuang, Xuya Wang, Michael Shen, Kun Yang, Giovanni Stracquadanio, Sarah M. Richardson, Yicong Lin, Lihui Wang, Roy Walker, Yisha Luo, Ping Sheng Ma, Huanming Yang, Yizhi Cai, Junbiao Dai, Joel S. Bader, Jef D. Boeke, Ying Jin Yuan

Research output: Contribution to journalArticlepeer-review


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 FIP1, and a loxPsym site affecting promoter function of ATP2. PoPM is a powerful tool for synthetic yeast genome debugging and an efficient strategy for phenotype-genotype mapping.

Original languageEnglish (US)
Article numbereaaf4706
Issue number6329
StatePublished - Mar 10 2017

ASJC Scopus subject areas

  • General


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