News

The Bacillus Genetic Stock Center is pleased to offer 16 new vectors designed for constructing fluorescent protein fusions. Three of the vectors come from the laboratory of Wolfgang Schumann at the University of Bayreuth, Germany, while the remaining 13 come from Peter Lewis at the University of Newcastle, Australia.

Sixteen new Fluorescent Protein tagging vectors

Jan van der Ploeg at the University of Zurich has kindly donated four Bacillus subtilis mutants with knockouts in one or more sulfur utilization genes: ssuA, ssuC, ssuD, cysI, or cysJ.

New Bacillus subtilis sulfur source utilization gene knockout mutants

From F. Denizot at the INRS in Marseille, France, comes pDG148-Stu a shuttle vector, capable of replicating in E. coli from the pBR322 origin and in Bacillus from the pUB110 origin. It allows inducible expression of foreign inserts cloned into its unique StuI site. Oriented, ligation-independent cloning of PCR fragments is possible using the proper primers and a prepared template. Erratum - The original Adobe Acrobat file describing pDG148-Stu was in error. Instead of treating StuI-linearized vector with T4 polymerase in the presence of dATP, one must use dTTP. The file above has been corrected.

Gram-Positive - E. coli Shuttle Vector Featuring Ligation-Independent Cloning and Inducible Expression

New from the Patrick Piggot lab at Temple University School of Medicine: a Bacillus subtilis integration vector that inserts into the dif site at about 166° on the chromosome. Integration takes place via the host system for resolving chromosome dimers and does not require the action of the standard recombination pathways. Even recA mutants can be transformed with this vector!

RecA-independent Integration Vector for Bacillus subtilis

A high titer PBS1 transducing lysate is now available, a gift from Brooke Murphy of the Tina Henkin lab here at the Ohio State University. The BGSC accession number for PBS1 is 1P1. Please note that this phage is heat labile and requires a motile strain of Bacillus subtilis as a host.

Vasant K. Chary from the Patrick Piggot lab at Temple University School of Medicine has kindly donated a pair of novel antibiotic-cassette switching vectors, pVK71 and pVK73, for use in Bacillus subtilis and other Gram-positive organisms.

Antibiotic Switching Vectors

The Bacillus Genetic Stock Center is pleased to offer an isogenic set of Bacillus subtilis mutants deficient in the major autolysins. Philippe Margot, from the Dimitri Karamata group at the Institut de Génétique et de Biologie Microbiennes in Lausanne, Switzerland, kindly donated the collection.

Bacillus subtilis autolysin-deficient mutants

From the laboratory of Samuel Singer, who retired in 1997 from Western Illinois University, comes a collection of environmental bacterial isolates demonstrating activity against a variety of invertebrate species.

Novel Strains Showing Insecticidal, Nematicidal, and Molluscicidal Activity

From the laboratory of Wolfgang Schumann at the University of Bayreuth come two new expression vectors capable of integrating into the Bacillus subtilis chromosome at the lacA locus. Each allows for regulated expression of cloned inserts.

Bacillus subtilis Integration Vectors with Inducible Expression of Cloned Inserts

pMUTIN4 should allow the researcher to produce knockout or conditional expression mutations in any unknown coding sequence.

A Vector Useful for Gram-Positive Genomics

From the laboratory of Chet Price at UC Davis comes an important set of mutants altered with the regulation of late stationary phase events. PrpC is a member of the PPM family of serine/threonine protein phosphatases; it removes phosphates from PrkC, a serine/threonine kinase. The pair is believed to regulate the activity of Elongation factor G (EF-G) during stationary phase. In prpC knockouts, such as PB702 (=BGSC 1A961), stationary phase cultures grow to a much greater density in rich, non-sporulation media. In contrast, stationary phase cultures of prkC knockouts, such as PB705 (=BGSC 1A962), grow to a significantly lower density. In double mutants, such as PB722 (=BGSC 1A964), prkC is epistatic to prpC (1). We thank Tatiana Gaidenko and Chet Price for donating this set of strains to the BGSC.

Neil Welker has donated plasmid pNW33N, a fifth generation vector that stably replicates in Bacillus subtilis, Geobacillus stearothermophilus and Escherichia coli

Cloning Vector for Thermophilic Bacillus Strains