Plant codon optimized cry genes of Bacillus thuringiensis can be expressed as soluble proteins in Escherichia coli BL21 Codon Plus strain as NusA-Cry protein fusions.
For optimal expression of delta-endotoxins from Bacillus thuringiensis in plants, preferential changes in the codon sequences, and reduction in overall AT content in the nucleotide sequence of the genes is important. Reports suggest that sequences with such modifications cannot be overexpressed in bacteria. We report here that the modified genes can be overexpressed in a strain of Escherichia coli carrying extra tRNA genes for some of the codons occurring at high frequency in plant genes and less preferred in E. coli. We also demonstrate that proteins when expressed as fusion products with NusA protein, are obtained as soluble fraction rather than in inclusion bodies. This allows easy and accurate LC50 analysis on insect pests.
Kumar
S
S
Department of Genetics, University of Delhi South Campus, Benito Juarez Road, New Delhi 110021, India.
Birah
A
A
Chaudhary
B
B
Burma
P K
PK
Gupta
G P
GP
Pental
D
D
D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics*; D003062:Codon; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D015151:Immunoblotting; D010445:Peptide Elongation Factors / Q000235:genetics*; D010572:Pest Control, Biological / Q000379:methods; D010944:Plants / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D035602:Transcriptional Elongation Factors
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D003062:Codon; D029968:Escherichia coli Proteins; D010445:Peptide Elongation Factors; D011994:Recombinant Proteins; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; C068727:nusA protein, E coli