광활성 메티오닌 표지단백질 생합성을 위한 메티오닐 tRNA 합성효소 및 이를 이용한 광활성 단백질 G 변이체 제조방법

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Title
광활성 메티오닌 표지단백질 생합성을 위한 메티오닐 tRNA 합성효소 및 이를 이용한 광활성 단백질 G 변이체 제조방법
Alternative Title
Methionyl tRNA synthase mutants for photoactivatable methionine mimetics and preparation method of photoactivatable protein G mutants
Inventor(s)
이명규; 이가비; 문정희; 정봉현
Assignee(s)
한국생명공학연구원
Application No.
14/894,756 (2015.11.30)
Registration No.
9914757 (2018.03.13)
Country
미국
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.
Type
Patent
Appears in Collections:
Division of Biomaterials Research > Bionanotechnology Research Center > 2. Patents
Division of Bio Technology Innovation > Core Facility Management Center > 2. Patents
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