Abstract
We describe a method, DNA array to protein array (DAPA), which allows the 'printing' of replicate protein arrays directly from a DNA array template using cell-free protein synthesis. At least 20 copies of a protein array can be obtained from a single DNA array. DAPA eliminates the need for separate protein expression, purification and spotting, and also overcomes the problem of long-term functional storage of surface-bound proteins.
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Acknowledgements
We thank H. Liu for expert technical assistance, B. Korn (DKFZ, Heidelberg) and J. Taipele (University of Helsinki) for providing clones. Research at the Babraham Institute is supported by the Biotechnology and Biological Sciences Research Council. Work on this project has been supported through the European Commission 6th Framework Programme Integrated Project MolTools.
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M.H. and M.J.T. devised the principle of DAPA; M.H. developed and exemplified the method; O.S. optimized the method, devised the DAPA apparatus and designed and performed the experiments shown in Figures 1 and 2; E.A.P. performed initial DAPA experiments, contributing examples to the data shown in Supplementary Figures 3 and 4; F.K. characterized the double-6His tag for protein immobilization and protein detection methods; O.E. spotted the DNA template slides for experiments shown in Figure 1; M.H., O.S. and M.J.T. prepared the manuscript.
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Supplementary Figures 1–4, Supplementary Methods (PDF 379 kb)
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He, M., Stoevesandt, O., Palmer, E. et al. Printing protein arrays from DNA arrays. Nat Methods 5, 175–177 (2008). https://doi.org/10.1038/nmeth.1178
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DOI: https://doi.org/10.1038/nmeth.1178
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