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| Expression and Purification of Enhanced Green Fluorescent Protein Based on Baculovirus Expression System |
| ZHANG Mei1, FU Yuan-hui2, HE Jin-sheng1,2, LU Yan-yan1, ZHENG Xian-xian1 |
1. Department of Immunology,Anhui Medical University, Hefei 230032, China;
2. College of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing 100044, China |
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Abstract Enhanced green fluorescent protein (EGFP) gene sequence with 5' end His tag was amplified by PCR. The recombinant baculovirus encoding EGFP was constructed and transfected into sf9 cells by Cellfectine reagent. The resultant fluorescence was observed by fluorescent microscope, and the expressed EGFP protein was purified by Ni column affinity chromatography and exploited as antigen to analyze specific humour immune response. The open reading frame (ORF) of EGFP with 5' end His tag was cloned successfully. The expression of EGFP from recombinant baculovirus was confirmed by fluorescent microscope, and the purified EGFP reached 90% purity and had the ability to bind specific antibody. The high efficient expression, simple protocol available for purification and intact biological activity are the advantages to prepare EGFP from baculovirus expression system.
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Received: 19 November 2010
Published: 27 May 2011
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