Invention Grant
- Patent Title: Methionyl tRNA synthetase for biosynthesis of photomethionine-labeled protein and method for preparing photoactive protein G variant using same
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Application No.: US14894756Application Date: 2014-05-29
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Publication No.: US09914757B2Publication Date: 2018-03-13
- Inventor: Myung Kyu Lee , Bong Hyun Chung , Jeong Hee Moon , Ga Bi Lee
- Applicant: Korea Research Institute of Bioscience and Biotechnology
- Applicant Address: KR Daejeon
- Assignee: Korean Research Institute of Bioscience and Biotechnology
- Current Assignee: Korean Research Institute of Bioscience and Biotechnology
- Current Assignee Address: KR Daejeon
- Agency: Banner & Witcoff, Ltd.
- Priority: KR10-2013-0061895 20130530; KR10-2013-0148287 20131202
- International Application: PCT/KR2014/004803 WO 20140529
- International Announcement: WO2014/193176 WO 20141204
- Main IPC: C07K14/315
- IPC: C07K14/315 ; C07K16/28 ; C12P21/00 ; C12N9/00

Abstract:
Provided is a methionyl tRNA synthase (MRS) for the biosynthesis of a photomethionine-labeled protein and a method for preparing a photoactivatable protein G variant using same and, more particularly, to an MRS variant in which alanine at the position of 12th is substituted with glycine, leucine at the position of 13th by serine, tyrosine at the position of 260th by phenylalanine, isoleucine at the position of 297th by valine, and histidine at the position of 301st by leucine from the N-terminal of the amino acid sequence of a wild-type Escherichia coli methionyl tRNA synthase. The MRS variant effectively confirms the biosynthesis of a photomethionine (pM)-labeled target protein and thus can be utilized for the biosynthesis of a pM-labeled target protein. In addition, a pM-introduced protein G variant, in which a plasmid encoding the MRS variant (MRS5m) and a PG-C3 plasmid, in which, in the third immunoglobulin G binding region of protein G, positions of 32nd, 35th, and 40th are substituted with a methionine (Met) residue and a position of 37th by an arginine (Arg) residue, are introduced into Escherichia coli and then refined, has a specific covalent bond with an antibody when subject to UV irradiation, and thus the pM-introduced protein G variant using the MRS variant can be utilized for producing a highly sensitive biochip, biosensor, or cell-capturing chip.
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