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[Special Issue Research Article] Bug mapping and fitness testing of chemically synthesized chromosome X

ScienceNOW - Fetched: March 9th, 2017, 3:00pm UTC

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. Authors: 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 Jia, Wen-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

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