Restriction digests of the clone give the following sizes (kb): EcoRI/BglI--5.0, 2.2, 1.1, 0.8; BamHI--9.4; BglI/BglII--3.5, 2.2, 2.0, 1.3. Structural stability of the plasmid in B. subtilis can be affected by high levels of protein production. Under these conditions, cell growth and stability may be improved by reducing the antibiotic concentration in the media. May not be suitable for cloning very strong expression signals. Translational fusion shuttle vector using 'lacZ as reporter. Also useful for the construction of fusion proteins. Neo confers resistance to neomycin and kanamycin and ble confers resistance to bleomycin and phleomycin. The 5' truncated 'lacZ gene, lacking transcriptional and translational initiation signals, is useful in both E. coli and in B. subtilis, since the endogenous beta-galactosidase level in B. subtilis is low. The order of the major features in the plasmid is: To terminator - HindIII/MCS/BamHI - 'lacZ - rrnB terminator - ampR - pMB1 ori - pUB110 ori - neoR - bleR. |
