James D Watson Institute Of Genome Science

Hangzhou, China

James D Watson Institute Of Genome Science

Hangzhou, China
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Zhang W.,Tsinghua University | Zhao G.,Tsinghua University | Luo Z.,Tsinghua University | Lin Y.,Tsinghua University | And 43 more authors.
Science | Year: 2017

We designed and synthesized a 976,067-base pair linear chromosome, synXII, based on native chromosome XII in Saccharomyces cerevisiae. SynXII was assembled using a two-step method, specified by successive megachunk integration and meiotic recombination-mediated assembly, producing a functional chromosome in S. cerevisiae. Minor growth defect "bugs" detected in synXII, caused by deletion of tRNA genes, were rescued by introducing an ectopic copy of a single tRNA gene. The ribosomal gene cluster (rDNA) on synXII was left intact during the assembly process and subsequently replaced by a modified rDNA unit used to regenerate rDNA at three distinct chromosomal locations. The signature sequences within rDNA, which can be used to determine species identity, were swapped to generate a Saccharomyces synXII strain that would be identified as Saccharomyces bayanus by standard DNA barcoding procedures. © 2017, American Association for the Advancement of Science. All rights reserved.


Wu Y.,Tianjin University | Li B.-Z.,Tianjin University | Zhao M.,Tianjin University | Mitchell L.A.,NYU Langone Medical Center | And 44 more authors.
Science | Year: 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. © 2017, American Association for the Advancement of Science. All rights reserved.

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