[Comparative study of the expression of the native and modified cry3a genes of Bacillus thuringiensis in prokaryotic and eukaryotic cells].
The cry3a gene of Bacillus thuriengiensis was cloned. Based on sequence analysis of this gene, a modified gene, cry3aM, was constructed, which has the optimal codon composition for effective expression in eukaryotic cells. Hybrid genes cry3a-licBM2 and cry3aM-licBM2 were constructed, in which the sequences of the native and modified genes are fusedfused with the reorter gene for thermostable lichenase in the reading frame. We have shown that the expression levels of hybrid genes cry3a-licBM2 and cry3aM-licBM2 in Escherichia coli are comparable, being 5% of those for reporter gene licBM2. In cells of a lower eukaryote Saccharomyces cerevisiae, the expression of hybrid gene cry3aM-licBM2? Which contains the modified gene, considerably exceeded the level of expression of cry3a-licBM2 containing the native gene. The presence of lichenase in the composition of hybrid proteins was shown to facilitate selection and analysis of the expression level of hybrid proteins in transgenic organisms.
Salekhi Dzhuzani
G R
GR
Komakhin
R A
RA
Piruzian
E S
ES
D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D048013:Acetivibrio thermocellus / Q000201:enzymology; D004731:Endotoxins / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005798:Genes, Bacterial; D017930:Genes, Reporter; D006026:Glycoside Hydrolases / Q000096:biosynthesis* / Q000235:genetics; D006460:Hemolysin Proteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D011993:Recombinant Fusion Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D006026:Glycoside Hydrolases; C028020:licheninase