International Journal of Industrial Entomology and Biomaterials

Korean Society of Sericultural Science (한국잠사학회)
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Expression of a Recombinant Bacillus thuringiensis $\delta$-Endotoxin Fused with Enhanced Green Fluorescent Protein in Escherichia coli

  • Je, Yeon-Ho (School of Agricultural Biotechnology, Seoul National University) ;
  • Roh, Jong-Yul (School of Agricultural Biotechnology, Seoul National University) ;
  • Li, Ming-Shun (School of Agricultural Biotechnology, Seoul National University) ;
  • Chang, Jin-Hee (School of Agricultural Biotechnology, Seoul National University) ;
  • Shim, Hee-Jin (School of Agricultural Biotechnology, Seoul National University) ;
  • Jin, Byung-Rae (College of National Resources and Life Science, Dong-A University) ;
  • Boo, Kyung-Saeng (School of Agricultural Biotechnology, Seoul National University)
  • Published : 2004.06.01
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Abstract

The expression of a fusion protein comprised of the B. thuringiensis crystal protein, Cry1Ac, and enhanced green fluorescent protein (EGFP) in Escherichia coli XLl-blue was examined. Three recombinant plasmids were transformed into E. coli XL1-blue and named as ProAc/Ec, MuEGFP/Ec and ProMu-EGFP/Ec, respectively. All transformants were observed by light and fluorescence microscopy at mid-log phase. The expression in E. coli transformants, ProMu-EGFP/Ec and MuEGFP/Ec, exhibited bright enough fluorescence to be observed. Furthermore, ProMu-EGFP/Ec produced fluorescent inclusions, which may have been recombinant crystals between EGFP and Cry1Ac while MuEGFP/Ec expressed soluble EGFP in cell. In SDS-PAGE, ProAc/Ec had 130 kDa crystal protein band and MuEGFP/Ec had thick 27 kDa EGFP band. However, ProMu-EGFP/Ec had about 150 kDa fusion protein band. Accordingly, these results indicated that a fusion protein between the B. thuringiensis crystal protein and a foreign protein under the lacZ promoter was successfully expressed as granular structure in E. coli. It is suggested that the E. coli expression system by N-terminal fusion of B. thuringiensis crystal protein may be useful as excellent means for fusion expression and characterization of B. thuringiensis fusion crystal protein.

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References

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