New shuttle vectors for ectopic insertion of genes into Bacillus subtilis.
We have constructed shuttle vectors for integration of genes via double homologous recombination into three ectopic sites on the chromosome of Bacillus subtilis. The sites of integration are the pyrD, gltA, and sacA genes located at 139 degrees, 172 degrees, and 333 degrees, respectively, on the chromosome. Integration of the vectors into the target genes leads to antibiotic resistance as well as different metabolic phenotypes. B. subtilis strains with integrations of the empty vectors were able to sporulate at rates comparable to wild type cells. Similar levels of expression were obtained from constitutive lacZ fusions integrated at the different sites.
Middleton
Rebecca
R
Department of Plant and Microbial Biology, University of California, Berkeley, USA. rebeccam@nature.berkeley.edu
Hofmeister
Antje
A
D001412:Bacillus subtilis / Q000235:genetics*; D001483:Base Sequence; D002876:Chromosomes, Bacterial / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D015870:Gene Expression; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic / Q000235:genetics*
D017931:DNA Primers