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Published on: June 23, 2016
Biotin- and Chloroalkane-Modified Nucleotides for Dual Display of Proteins on DNA Through Streptavidin and Halotag
Giammarco Liuni1,2, Lenka Poštová Slavětínská1, Miroslav Hájek1
1Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Prague, Czech Republic.
Abstract:
We designed and synthesized 2'-deoxyribonucleoside triphosphates (dNTPs) bearing chlorohexyl and biotin modifications attached to position 7 of 7-deazaadenine or to position 5 of cytosine. The modified dNTPs served as substrates for KOD XL DNA polymerase in the enzymatic construction of DNA bearing either one or a combination of the two modifications. The chlorohexyl-linked DNA underwent covalent reactions with halotag protein, while the biotin-linked DNA formed stable non-covalent complexes with monomeric streptavidin. The dually modified DNA bearing both modifications at different distances apart from each other were used for sequence-specific, spatially defined display of both halotag and streptavidin proteins. The study provides the foundation for display of proteins on DNA rulers.
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