caa a22 4500 465823963 CHVBK 20180323112143.0 cr unu---uuuuu 170327e19900901xx s 000 0 eng 10.1007/BF00604933 doi (NATIONALLICENCE)springer-10.1007/BF00604933 Regulated expression of heterologous genes in Bacillus subtilis using the Tn 10 encoded tet regulatory elements [Elektronische Daten] [Manfred Geissendörfer, Wolfgang Hillen] Summary: TheEscherichia coli-derivedtet regulatory elements from Tn10 have been used to construct vectors allowing the regulated, inducible, high-level expression of foreign genes inBacillus subtilis. While the wild-typetet promoters are inactive inB. subtilis, a synthetic mutanttet sequence with improved promoter consensus sequences and upstream poly A blocks shows activity inB. subtilis. The expression of an indicatorcat gene is inducible by sublethal amounts of tetracycline, indicating that the Tet repressor protein and thetet operator sequences are functional. However, the inducibility and maximal expression are not sufficient in this construct. To improve these properties atet operator sequence was placed between the —35 and —10 boxes of theB. subtilis-derived very strongxyl promoter. In the presence of atetR gene this construct is about 100-fold inducible and has high promoter strength, but some basal expression. This is avoided by placing a secondtet operator downstream resulting in no detectable basal expression at the expense of reduced inducibility. Using the system with a singletet operator inducible expression of glucose dehydrogenase fromB. megaterium was obtained at a very high level, and inducible expression of human single-chain urokinase-like plasminogen activator was achieved at the same level as inE. coli. Unlike inE. coli, the product was not degraded up to 4 h after induction inB. subtilis. These results demonstrate that the regulated expression vector described here should be very useful for production of foreign gene products fromB. subtilis cultures. Springer-Verlag, 1990 Geissendörfer Manfred Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie und Biochemie, Friedrich-Alexander Universität Erlangen-Nürnberg, Staudtstrasse 5, D-8520, Erlangen, Federal Republic of Germany aut Hillen Wolfgang Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie und Biochemie, Friedrich-Alexander Universität Erlangen-Nürnberg, Staudtstrasse 5, D-8520, Erlangen, Federal Republic of Germany aut Applied Microbiology and Biotechnology Springer-Verlag 33/6(1990-09-01), 657-663 0175-7598 33:6<657 1990 33 253 https://doi.org/10.1007/BF00604933 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00604933 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Geissendörfer Manfred Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie und Biochemie, Friedrich-Alexander Universität Erlangen-Nürnberg, Staudtstrasse 5, D-8520, Erlangen, Federal Republic of Germany aut NATIONALLICENCE 700 1- Hillen Wolfgang Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie und Biochemie, Friedrich-Alexander Universität Erlangen-Nürnberg, Staudtstrasse 5, D-8520, Erlangen, Federal Republic of Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer-Verlag 33/6(1990-09-01), 657-663 0175-7598 33:6<657 1990 33 253 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer