ISSN:
1617-4623
Keywords:
Key wordsBacillus subtilis
;
T7 RNA polymerase
;
Promoter specificity
;
Gene expression
;
Protein secretion
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract The adaptation and application of the Escherichia coli T7 RNA polymerase system for regulated and promoter-specific gene expression in Bacillus subtilis is reported. The expression cassette used in Bacillus subtilis was tightly regulated and T7 RNA polymerase (T7 RNAP) appeared 30 min after induction. The efficiency of T7 promoter-specific gene expression in B. subtilis was studied using one secretory and two cytosolic proteins of heterologous origin. The accumulation of E. coliβ-galactosidase, as well as a 1,4-β-glucosidase from Thermoanaerobacter brockii in B. subtilis after T7 RNAP induction was strongly enhanced by rifampicin inhibition of host RNAP activity. The α-amylase of Thermoactinomyces vulgaris, a secretory protein, was found to accumulate in the culture supernatant up to levels of about 70 mg/l 10–20 h after T7 RNAP induction, but was also deposited in cellular fractions. The addition of rifampicin inhibited α-amylase secretion, but unexpectedly, after a short period, also prevented its further (intra)cellular accumulation
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF02174183
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