Earticle

초록

영어

Here we describe a highly specific, robust and rapid new method for labeling of cell surface proteins with fluorescent probes. The method uses the trans-splicing reaction of Nostoc punctiforme (Npu) DnaE split-intein. Split-intein based protein trans-splicing reaction has been used for site-specific labeling of proteins in vitro for various applications. The trans-splicing reaction is self-processing reaction and can be used for introduction of various synthetic probes to target proteins without a large linker proteins. In this report, we have utilized split-intein based protein trans-splicing reaction for site specific labeling of cell surface proteins. We have prepared a model cell surface protein fused to N-terminal fragment of Npu DnaE Intein and C-terminal fragment of Npu DnaE Intein fused to a fluorescent probe. The model protein was expressed on the cell surface and the proteins were labeled with synthetic fluorescent probes through specific interaction between Nand C-fragment of inteins. This labeling reaction was performed within 30 mins using low concentration of fluorescent molecules (4 uM). No external energy was required for the labeling reaction. This improved labeling method will provide a useful tool for studying functions and mobility of proteins in live cells.

키워드

저자

• Kyoungmi MIN [ Dept. of medical engineering, Doungguk Unibersity, Seoul, 100-715. ]
• Ducho JUNG [ Dept. of medical engineering, Doungguk Unibersity, Seoul, 100-715. ]
• Youngeun KWON [ Dept. of medical engineering, Doungguk Unibersity, Seoul, 100-715. ]
• Jaejung LEE [ Dept. of medical engineering, Doungguk Unibersity, Seoul, 100-715. ]
• Youngtae CHANG [ Dept. of medical engineering, Doungguk Unibersity, Seoul, 100-715. ]

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간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576