Restriction digests of the clone give the following sizes (kb): BamHI--5.7, EcoRI--5.7, PstI--4.9, 0.7. Cloning into the NdeI, NheI, BamHI, SmaI or EcoRI sites inactivates lacZalpha. The NcoI site interrupts the chloramphenicol resistance sequence. This is the preferred strain for isolating the plasmid, because the copy number is higher in E. coli than in B. subtilis. Permits alpha-complementation to identify recombinants when used with Bacillus subtilis 6GM15, and alpha-complementation with plasmid marker rescue when used with Bacillis subtilis 6GM15[pHPS9R] (ATCC 37818). The cat86::lacZalpha fusion is in-frame. Expression is controlled by the P59 promoter from Lactococcus lactis subsp. cremoris Wg2. |
