Monday, June 27, 2016

Efficient Formation of Site-Specific Protein–DNA Hybrids Using Copper-Free Click Chemistry

Ann Mukhortava and Michael Schlierf

Protein–DNA hybrids have become increasingly popular molecular building blocks in bionanotechnology and single-molecule studies to synergistically combine the programmability of DNA with the chemical diversity of proteins. The growing demand for protein–DNA hybrids requires powerful strategies for their conjugation. Here, we present an efficient two-step method for protein–DNA assembly based on copper-free click chemistry. The method allows site-specificity and high coupling efficiency, while maintaining the conservation of protein activity. We compare our method to a commonly used protocol of direct linkage of maleimide-modified oligos. We demonstrate the significantly higher yield with a protein–DNA conjugate, which is analyzed using single-molecule force spectroscopy.

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