Construction of Improved Bacteriophage 105 Vectors for Cloning by Transfection in Bacillus subtilis

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Abstract
A series of improved phage vectors have been constructed, based on Bacillus subtilis bacteriophage .vphi.105, which can be used to clone gens in B. subtilis by direct transfection of protoplasts. The new vectors, designated .vphi.105J23, .vphi.105J24, .vphi.105J27 and .vphi.105J28, show frequencies of plaque formation that are equal to those of wild-type .vphi.105. This represents at least a 10-fold improvement over .vphi.105J9, the vector used in previous cloning experiments. Two of the new vectors .vphi.105J27 and .vphi.105J28 incorporate a mutation, cts-52, the renders that prophage temperature inducible. This has made it possible to devise a rapid small-scale procedure for screening progeny phage for the presence of inserted DNA. The usefulness of the new vectors in illustrated in the accompanying paper by cloning more than 20 B. subtilis sporulation genes.