Restriction digests of the clone give the following sizes (kb): EcoRI--6.7; BamHI--6.7; BglI/BglII--3.5, 3.2. 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. Promoter-cloning shuttle vector using the expression of chloramphenicol acetyltransferase (CAT) as the reporter. Also useful for construction of fusion proteins. Neo confers resistance to neomycin and kanamycin and ble confers resistance to bleomycin and phleomycin. The cat gene was derived from pUB112 by deletion of the promoter and the regulatory inverted repeat, resulting in constitutive cat gene expression when provided with an appropriate upstream promoter. The order of the major features in the plasmid is: To terminator - HindIII/MCS/EcoRI - cat - rrnB terminator - ampR - pMB1 ori - pUB110 ori - neoR - bleR. |
