Simple Purification of a Foreign Protein Using Polyhedrin Fusion in a Baculovirus Expression System
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概要
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Previously, we found that expression by translational fusion of the polyhedrin (Polh)-green fluorescence protein (GFP) led to the formation of granular structures, and that these fluorescent granules were easily precipitated by high-speed centrifugation. Here, we developed an easy, fast, mass purification system using this baculovirus expression system (BES). An enhanced GFP (EGFP) fused with the Polh gene at the N-terminus, including a linker and enterokinase (EK) site between Polh and EGFP, was expressed in Sf9 cells. The cells infected by AcPolhEKA-EGFP produced fluorescent granules. The EGFP fusion protein was purified from granule-containing cells in three steps: cell harvest, sonication, and EK digestion. Through final enterokinase digestion, EGFP presented mainly in the supernatant, and this supernatant fraction also showed a pure EGFP band. These results suggest that a combined procedure of Polh fusion expression and enterokinase digestion can be used for rapid and easy purification of other proteins.
- 2010-08-23
著者
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Roh Jong
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National U
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Choi Jae
Research Institute for Agriculture and Life Sciences, Seoul National University
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Kang Joong
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National U
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WANG Yong
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National U
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SHIM Hee
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National U
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LIU Qin
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National U
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TAO Xueying
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National U
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XU Hong
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National U
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HYUN Jin-Ho
Department of Biosystems and Biomaterials Science and Engineering, College of Agriculture and Life S
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WOO Soo
Department of Plant Medicine, College of Agriculture, Life and Environment Sciences, Chungbuk Nation
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JIN Byung
College of Natural Resources and Life Science, Dong-A University
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JE Yeon
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National U
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Jin Byung
College Of Natural Resources And Life Science Dong-a University
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Shim Hee
Department Of Agricultural Biotechnology College Of Agriculture And Life Sciences Seoul National Uni
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Tao Xueying
Department Of Agricultural Biotechnology College Of Agriculture And Life Sciences Seoul National Uni
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Hyun Jin-ho
Department Of Biosystems And Biomaterials Science And Engineering College Of Agriculture And Life Sc
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Choi Jae
Research Institute For Agriculture And Life Sciences Seoul National University
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Woo Soo
Department Of Plant Medicine College Of Agriculture Life And Environment Sciences Chungbuk National
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Xu Hong
Department Of Agricultural Biotechnology College Of Agriculture And Life Sciences Seoul National Uni
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KANO Joong
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National U
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Roh Jong
Department Of Agricultural Biotechnology College Of Agriculture And Life Sciences Seoul National Uni
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Kano Joong
Department Of Agricultural Biotechnology College Of Agriculture And Life Sciences Seoul National Uni
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Liu Qin
Department Of Agricultural Biotechnology College Of Agriculture And Life Sciences Seoul National Uni
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Je Yeon
Department Of Agricultural Biotechnology College Of Agriculture And Life Sciences Seoul National Uni
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Wang Yong
Department Of Agricultural Biotechnology College Of Agriculture And Life Sciences Seoul National Uni
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