Journal
SCIENCE
Volume 355, Issue 6329, Pages -Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.aaf4706
Keywords
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Categories
Funding
- 863 program [2012AA02A708]
- 973 program [2014CB745100]
- Ministry of Science and Technology [2015DFA00960]
- China National Natural Science Foundation of China [21390203, 21621004]
- U.S. NSF [MCB-1026068, MCB-1158201, MCB-1445545]
- U.S. Department of Energy [DE-FG02097ER25308]
- Chancellor's Fellowship from the University of Edinburgh
- Scottish Universities Life Sciences Alliance (SULSA)
- Biotechnology and Biological Sciences Research Counci [BB/M005690/1, BB/M025640/1, BB/M00029X/1]
- Direct For Biological Sciences
- Div Of Molecular and Cellular Bioscience [1445545, 1616111] Funding Source: National Science Foundation
- Biotechnology and Biological Sciences Research Council [BB/M025640/1, BBS/OS/GC/000001, BB/M00029X/1, BB/M005690/1, BB/M005690/2] Funding Source: researchfish
- Engineering and Physical Sciences Research Council [EP/P017401/1] Funding Source: researchfish
- BBSRC [BB/M005690/1, BB/M005690/2, BB/M00029X/1, BB/M025640/1] Funding Source: UKRI
- EPSRC [EP/P017401/1] Funding Source: UKRI
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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 loxPsymsite affecting promoter function of ATP2. PoPM is a powerful tool for synthetic yeast genome debugging and an efficient strategy for phenotype-genotype mapping.
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