期刊
NATURE PROTOCOLS
卷 3, 期 3, 页码 357-362出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2007.538
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资金
- NCI NIH HHS [U19 CA113297] Funding Source: Medline
- NIAID NIH HHS [AI52218] Funding Source: Medline
- NATIONAL CANCER INSTITUTE [U19CA113297] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [R01AI052218] Funding Source: NIH RePORTER
This protocol details the application of a high-throughput fluorescence-based screen, in conjunction with error-prone PCR/saturation mutagenesis, for altering the proficiency and/or promiscuity of a secondary metabolite glycosyltransferase (GT) via directed evolution. Given the structural and mechanistic similarities among secondary metabolite-associated GTs, this approach may provide a template for engineering other members of the GT-B superfamily.
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