Human deoxycytidine kinase as a conditional mutator in Escherichia coli |
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| Affiliation: | 1. College of Pharmaceutical Science & Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang University of Technology, Hangzhou, China;2. College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, China;3. National Institute of Diabetes and Digestive and Kidney, Diseases (NIDDK), National Institutes of Health, Bethesda, MD, USA;4. Center for Human Nutrition, David Geffen School of Medicine, University of California, Los Angeles, CA, USA;5. Zhejiang Institute for Food and Drug Control, Hangzhou, China |
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| Abstract: | The chemical diversification of DNA precursors was undertaken in Escherichia coli by expressing the human gene for deoxycytidine kinase, and supplying such recombinant strains with nucleoside analogues bearing an altered base or sugar. Arabinocytidine and dideoxycytidine thus became highly toxic to E. coli in the sub-millimolar range. Deoxynucleosides bearing isoadénine (2-aminopurine) and isoguanine (2-hydroxy-6-aminopunne) showed a high mutagenic potency towards the recombinant strains, to an extent comparable to that of the most efficient mutator alleles (dnaQ). These findings open the way to the propagation of chemically remodelled nucleic acids and to the controlled hypermutagenesis of plasmids in vivo. |
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