2013 Fiscal Year Final Research Report

Creaction of artificial endonuclease using DNA-cleavage domain of a novel restriction enzyme

Research Project

Project/Area Number	24658077
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Single-year Grants
Research Field	Applied microbiology
Research Institution	Kyoto University
Principal Investigator	KITA Keiko 京都大学, (連合)農学研究科(研究院), 教授 (70234226)
Research Collaborator	NAKAMURA Keigo 京都大学, 農学部, 4回生 HIOKI Takahiro 京都大学, 農学部, 4回生
Project Period (FY)	2012-04-01 – 2014-03-31
Keywords	制限酵素 / 分子工学 / DNA / ドメイン / リンカー / 分子間相互作用 / 構造機能相関 / 人工酵素
Research Abstract	Restriction enzymes recognize and cleave DNA at specific site, and are both indispensable tools for gene manipulation and for studying sequence-specific interaction with DNA. Co-crystal structure of EcoT38I reveals several unique features for DNA recognition and catalysis. This study demonstrates that site-directed mutagenesis approach yielded a EcoT38I (GPuGCPyC) variant that preferentially cleaves one of the degenerate sequences (GAGCTC).