Amplification of the Escherichia coli lacZ gene in Bacillus subtilis and its expression on a by-product growth medium.
A Bacillus subtilis wild type strain and a kinA (spoIIJ) isogenic mutant were compared as hosts for the expression of the Escherichia coli beta-galactosidase gene, lacZ, driven by the B. subtilis aprE promoter in a chromosomal system. The 2 x SG sporulation formula, with some modifications, was used as a basal medium. The specific activity values recorded by the mutant strain at the stationary phase were markedly higher than those achieved by the wild type host. Exposure of the cells to increasing levels of chloramphenicol resulted in significant amplifications of the lacZ region. Gene copy numbers of 19 and 11 were estimated in the amplified wild type and kinA strains, respectively, with high segregational stability records. The magnitude of beta-galactosidase over-expression was dependent on, and roughly proportional to antibiotic resistance levels. Among five examined by-products, a 2.3-times diluted concentration of neutralized cheese whey was successfully used as a sole medium component for over-expression of the recombinant beta-galactosidase gene in B. subtilis.
el-Helow
E R
ER
Department of Botany, Faculty of Science, University of Alexandria, Alexandria, Egypt. elhelow@uclink4.berkeley.edu
Ghanem
K M
KM
Mohamad
E A
EA
D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D003470:Culture Media; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005784:Gene Amplification; D018628:Gene Dosage; D005798:Genes, Bacterial; D007763:Lac Operon; D011994:Recombinant Proteins / Q000096:biosynthesis*; D001616:beta-Galactosidase / Q000096:biosynthesis* / Q000235:genetics
D003470:Culture Media; D011994:Recombinant Proteins; D001616:beta-Galactosidase