Bug mapping and fitness testing of chemically synthesized chromosome X.
Yi WuBing-Zhi LiMeng ZhaoLeslie A MitchellZe-Xiong XieQiu-Hui LinXia WangWen-Hai XiaoYing WangXiao ZhouHong LiuXia LiMing-Zhu DingDuo LiuLu ZhangBao-Li LiuXiao-Le WuFei-Fei LiXiu-Tao DongBin JiaWen-Zheng ZhangGuo-Zhen JiangYue LiuXue BaiTian-Qing SongYan ChenSi-Jie ZhouRui-Ying ZhuFeng GaoZheng KuangXuya WangMichael ShenKun YangGiovanni StracquadanioSarah M RichardsonYicong LinLihui WangRoy S K WalkerYisha LuoPing-Sheng MaHuanming YangYizhi CaiJun-Biao DaiJoel S BaderJef D BoekeYing-Jin YuanPublished in: Science (New York, N.Y.) (2017)
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.