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Expression and characterization of a recombinant Cry1Ac crystal protein with enhanced green fluorescent protein in acrystalliferous Bacillus thuringiensis
Author(s) -
Roh J.Y.,
Li M.S.,
Chang J.H.,
Choi J.Y.,
Shim H.J.,
Shin S.C.,
Boo K.S.,
Je Y.H.
Publication year - 2004
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2004.01505.x
Subject(s) - cry1ac , bacillus thuringiensis , green fluorescent protein , fusion protein , microbiology and biotechnology , biology , recombinant dna , fusion gene , protein a/g , bipyramid , chemistry , gene , biochemistry , transgene , genetically modified crops , bacteria , crystal structure , crystallography , genetics
Summary Aims: To investigate fusion expression between Bacillus thuringiensis crystal protein and a foreign protein, the expression of a fusion protein comprised of Cry1Ac, and enhanced green fluorescent protein (EGFP) in B. thuringiensis Cry − B strain was examined. Methods and Results: The N‐terminal fusion expression of EGFP in Cry1Ac was attempted under the control of the native cry1Ac promoter. The EGFP gene was cloned into pProMu and named pProMu‐EGFP. The transformant, ProMu‐EGFP/CB produced parasporal inclusions that were of bipyramidal‐shaped crystals in size ranging from 200 to 300 nm. The fusion protein was approximately 150 kDa and identified by the immunoblot analysis using a Cry1Ac antibody and also a GFP antibody. The LC 50 of the ProMu‐EGFP/CB was twofold higher when compared with that by the ProAc/CB. However, the crystal protein produced by the ProMu‐EGFP/CB was effective on Plutella xylostella larvae. Conclusions: The ProMu‐EGFP/CB produced bipyramidal shaped and insecticidal crystals comprising fusion proteins. Significance and Impact of the Study: Through the N‐terminal fusion expression of EGFP and Cry1Ac, expression and crystallization between the B. thuringiensis crystal protein and a foreign protein were validated.