New shuttle vector for cloning in Bacillus stearothermophilus.
Cloning vector plasmid pRP9 was constructed on the basis of the broad host-range plasmid pLM6. pRP9 was a small plasmid (2.9 kb), possessed a convenient polyrestriction site sequence and efficiently transformed Bacillus subtilis, Bacillus stearothermophilus and Escherichia coli. Furthermore, pRP9 presented a very high segregational stability in Bacillus hosts. Also, the structural stability in Bacillus strains, grown under selective pressure, of pRP9 carrying a 3-kb fragment, was high. No single-stranded and high-molecular weight pRP9 DNA was found in B. stearothermophilus. The host/vector systems described possessed all the properties required for efficient gene cloning.
De Rossi
E
E
Department of Genetics and Microbiology A. Buzzati Traverso, University of Pavia, Italy.
Brigidi
P
P
Welker
N E
NE
Riccardi
G
G
Matteuzzi
D
D
D001412:Bacillus subtilis / Q000235:genetics / Q000254:growth & development; D003001:Cloning, Molecular / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development; D005822:Genetic Vectors / Q000235:genetics*; D001411:Geobacillus stearothermophilus / Q000235:genetics* / Q000254:growth & development; D066298:In Vitro Techniques; D010957:Plasmids / Q000235:genetics*
D004269:DNA, Bacterial