Open AccessEfficient Bacillus subtilis Cloning System using Bacteriophage Vector 01Q5J9
Author(s) -
Jeff Errington
Publication year - 1984
Publication title -
microbiology
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.019
H-Index - 179
eISSN - 1465-2080
pISSN - 1350-0872
DOI - 10.1099/00221287-130-10-2615
Subject(s) - bacillus subtilis , cloning vector , cloning (programming) , bacteriophage , biology , molecular cloning , recombinant dna , multiple cloning site , plasmid , escherichia coli , genomic library , gene , vector (molecular biology) , genetics , microbiology and biotechnology , computational biology , bacteria , gene expression , peptide sequence , computer science , programming language
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.