論文

査読有り
2017年6月

Design of Modular Protein Tags for Orthogonal Covalent Bond Formation at Specific DNA Sequences

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
  • Thang Minh Nguyen
  • ,
  • Eiji Nakata
  • ,
  • Masayuki Saimura
  • ,
  • Huyen Dinh
  • ,
  • Takashi Morii

139
25
開始ページ
8487
終了ページ
8496
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1021/jacs.7b01640
出版者・発行元
AMER CHEMICAL SOC

Simultaneous formation of specific covalent linkages at nucleotides in given DNA sequences demand distinct orthogonal reactivity of DNA modification agents. Such highly specific reactions require well-balanced reactivity and affinity of the DNA modification agents. Conjugation of a sequence-specific DNA binding zinc finger protein and a self-ligating protein tag provides a modular adaptor that expedites formation of a covalent bond between the protein tag and a substrate-modified nucleotide at a specific DNA sequence. The modular adaptor stably locates a protein of interest fused to it at the target position on DNA scaffold in its functional form. Modular adaptors with orthogonal selectivity and fast reaction kinetics to specific DNA sequences enable site-specific location of different protein molecules simultaneously. Three different modular adaptors consisting of zinc finger proteins with distinct DNA sequence specificities and self-ligating protein tags with different substrate specificities achieved orthogonal covalent bond formation at respective sequences on the same DNA scaffold with an overall coassembly yield over 90%. Application of this unique set of orthogonal modular adaptors enabled construction of a cascade reaction of three enzymes from xylose metabolic pathway on DNA scaffold.

リンク情報
DOI
https://doi.org/10.1021/jacs.7b01640
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000404809600023&DestApp=WOS_CPL
ID情報
  • DOI : 10.1021/jacs.7b01640
  • ISSN : 0002-7863
  • Web of Science ID : WOS:000404809600023

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