Abstract
We have expressed the pneumolysin gene of Streptococcus pneumoniae in Bacillus subtilis, both from its own promoter and as a fusion protein. The level of expression of pneumolysin from its own promoter was low. The protein produced was hemolytically active. A higher level of expression (about 10 micrograms/ml of culture) was achieved when either one of two C-terminal fragments (corresponding to amino acids 265-471 and 55-471, respectively) or the entire coding part of the pneumolysin gene were fused to the promoter and signal sequence-coding region of the alpha-amylase gene of Bacillus amyloliquefaciens. The C-terminal fusion peptides reacted with anti-pneumolysin serum, but were not hemolytically active. In both cases most of the peptide remained cell-associated. When the entire pneumolysin gene was fused to the signal sequence, a hemolytically active form of pneumolysin could be detected, and most of the product was found in a processed form in the culture supernatant. The full-length pneumolysin secreted from B. subtilis was partially purified and used as antigen in an enzyme immunoassay with rabbit anti-pneumolysin serum.
References
Jun 1, 1978·Molecular & General Genetics : MGG·J KreftW Goebel
Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·H TowbinJ Gordon
Mar 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·S D Ehrlich
Mar 1, 1986·Journal of General Microbiology·P KallioM Sarvas
Jan 1, 1987·The Journal of Infection·Y KerttulaP H Mäkelä
Jan 1, 1986·Pediatric Infectious Disease·M LeinonenP H Mäkelä
Aug 15, 1970·Nature·U K Laemmli
Jan 1, 1982·Molecular & General Genetics : MGG·C P MoranR Losick
Sep 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·I PalvaL Kääriäinen
May 1, 1983·Gene·I PalvaC Weissmann
Sep 1, 1983·Gene·R F PetterssonI Palva
Apr 21, 1984·British Medical Journal·M R Lockley, R Wise
Jan 1, 1982·Infection·L A Soares, C E Melles
Jul 1, 1983·The Journal of Infectious Diseases·R M DouglasD J Hansman
Citations
Nov 1, 1995·Clinical and Diagnostic Laboratory Immunology·A VirolainenM Leinonen
Nov 30, 1990·Gene·L JørgensenB Diderichsen
Oct 1, 1990·Current Opinion in Biotechnology·G Rapoport, A Klier
Sep 1, 1989·The Journal of Infection·E JalonenM Leinonen
Feb 1, 1996·The Pediatric Infectious Disease Journal·A VirolainenM Leinonen
Jan 15, 1992·European Journal of Biochemistry·J M Sanchez-PuellesE Garcia
Mar 18, 2000·Antimicrobial Agents and Chemotherapy·M J FerrándizA G De La Campa
Jun 25, 2003·Antimicrobial Agents and Chemotherapy·Luz BalsalobreAdela G de la Campa
Dec 8, 2005·Applied and Environmental Microbiology·Roger S BongersMichiel Kleerebezem
Dec 4, 2004·Journal of Bacteriology·Patricia RomeroErnesto García
Jul 22, 2005·Journal of Applied Microbiology·M VitikainenM Sarvas
Apr 28, 2004·Virology·Patricia RomeroErnesto García
Feb 5, 2005·Anais Da Academia Brasileira De Ciências·Luís C S FerreiraWolfgang Schumann
Nov 1, 1995·Acta Oto-laryngologica·A VirolainenJ Eskola
Jan 5, 2000·Clinical and Diagnostic Laboratory Immunology·J A ScottM Leinonen
Aug 15, 1994·FEMS Microbiology Letters·R H OwenT J Mitchell
Nov 18, 1998·Journal of Clinical Microbiology·A FenollJ Casal
Jul 1, 1993·Infection and Immunity·P SaloM Leinonen
Jul 1, 1995·Infection and Immunity·J KuoA A Lindberg
Mar 1, 1993·Microbiological Reviews·M Simonen, I Palva