A 20-kilodalton protein is required for efficient production of the Bacillus thuringiensis subsp. israelensis 27-kilodalton crystal protein in Escherichia coli
Autor: | L F Adams, J E Visick, H. R. Whiteley |
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Rok vydání: | 1989 |
Předmět: |
Bacterial Toxins
Molecular Sequence Data Restriction Mapping Bacillus thuringiensis Biology Molecular cloning medicine.disease_cause Microbiology Gene product Hemolysin Proteins DDB1 Bacterial Proteins HSPA2 Escherichia coli medicine Amino Acid Sequence Cloning Molecular Molecular Biology Gene Bacillus thuringiensis Toxins Base Sequence Molecular biology Endotoxins Molecular Weight Open reading frame Genes Genes Bacterial AKT1S1 Research Article |
Zdroj: | Journal of Bacteriology. 171:521-530 |
ISSN: | 1098-5530 0021-9193 |
Popis: | The 27-kilodalton (kDa) mosquitocidal protein gene from Bacillus thuringiensis subsp. israelensis has been cloned as a 10-kilobase (kb) HindIII fragment from plasmid DNA; efficient expression in Escherichia coli KM1 depends on a region of DNA located approximately 4 kb upstream (K. McLean and H. R. Whiteley, J. Bacteriol. 169:1017-1023, 1987). We have cloned the upstream DNA region and show that it contains a complete open reading frame (ORF) encoding a protein with a molecular mass of 19,584 Da. Sequencing of adjacent stretches of DNA revealed two partial ORFs: one has 55.2% identity in an overlap of 319 amino acids to the putative transposase of IS231 of B. thuringiensis subsp. thuringiensis, and the other, a 78-codon partial ORF, may be the carboxyl terminus of the 67-kDa protein previously observed in maxicells of strain KM1. A 0.8-kb fragment containing only the 20-kDa protein gene greatly enhanced the expression of the 27-kDa protein in E. coli. The introduction of nonsense codons into the 20-kDa protein gene ORF abolished this effect, indicating that the gene product, not the mRNA or DNA, is required for the enhancement. The effect of the 20-kDa protein gene on various fusions of lacZ to the 27-kDa protein gene suggests that the 20-kDa protein acts after the initiation of translation of the 27-kDa protein gene. |
Databáze: | OpenAIRE |
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