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