期刊
NATURE METHODS
卷 12, 期 3, 页码 203-+出版社
NATURE PORTFOLIO
DOI: 10.1038/NMETH.3223
关键词
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资金
- US National Science Foundation [DGE-0718124]
- US National Institutes of Health (NIH) Pioneer Award [DP1HG007811]
- NIH Biomedical Technology Research Resource project [P41GM103533]
- Howard Hughes Medical Institute
Random mutagenesis methods only partially cover the mutational space and are constrained by DNA synthesis length limitations. Here we demonstrate programmed allelic series (PALS), a single-volume, site-directed mutagenesis approach using microarray-programmed oligonucleotides. We created libraries including nearly every missense mutation as singleton events for the yeast transcription factor Gal4 (99.9% coverage) and human tumor suppressor p53 (93.5%). PALS-based comprehensive missense mutational scans may aid structure-function studies, protein engineering, and the interpretation of variants identified by clinical sequencing.
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