Evolution of a cytokine using DNA family shuffling
| Autor: | Phillip A. Patten, Teddy Chen, Glenn Dawes, Brett W. Cox, Juha Punnonen, Chia-Chun J. Chang, Willem P. C. Stemmer |
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| Rok vydání: | 1999 |
| Předmět: |
Models
Molecular Molecular Sequence Data Biomedical Engineering Clone (cell biology) Mutagenesis (molecular biology technique) Bioengineering CHO Cells Biology medicine.disease_cause Antiviral Agents Applied Microbiology and Biotechnology Cell Line Evolution Molecular Mice chemistry.chemical_compound Molecular evolution Cricetinae medicine Animals Humans Amino Acid Sequence Encephalomyocarditis virus Gene Genetics Mutation Sequence Homology Amino Acid Point mutation Interferon-alpha DNA DNA shuffling chemistry Mutagenesis Molecular Medicine Biotechnology |
| Zdroj: | Nature Biotechnology. 17:793-797 |
| ISSN: | 1546-1696 1087-0156 |
| Popis: | DNA shuffling of a family of over 20 human interferon-alpha (Hu-IFN-alpha) genes was used to derive variants with increased antiviral and antiproliferation activities in murine cells. A clone with 135,000-fold improved specific activity over Hu-IFN-alpha2a was obtained in the first cycle of shuffling. After a second cycle of selective shuffling, the most active clone was improved 285,000-fold relative to Hu-IFN-alpha2a and 185-fold relative to Hu-IFN-alpha1. Remarkably, the three most active clones were more active than the native murine IFN-alphas. These chimeras are derived from up to five parental genes but contained no random point mutations. These results demonstrate that diverse cytokine gene families can be used as starting material to rapidly evolve cytokines that are more active, or have superior selectivity profiles, than native cytokine genes. |
| Databáze: | OpenAIRE |
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