Efficient Bacillus subtilis cloning system using bacteriophage vector phi 105J9

Abstract

An efficient system for cloning in Bacillus subtilis is described which uses a newly constructed bacteriophage vector, phi 105J9. The phage genome contains cloning sites for the enzymes BamH1, XbaI and SalI, and can accommodate inserts of passenger DNA of at least 4 kbp. Recombinant phages, which can both plaque and lysogenize normally, are recovered after direct transfection of protoplasts in the presence of polyethylene glycol. Several fully functional sporulation genes and one biosynthetic gene from B. subtilis have been isolated from genomic libraries that were constructed with the new vector. The system may provide an alternative to some of the cloning methods currently available that use Escherichia coli as host.