Restriction digests of the clone give the following sizes (kb): EcoRI--6.8; BamHI--6.8; BglI/BglII--3.5, 3.3. The Bacillus subtilis promoter vegII initiates transcription in both B. subtilis and E. coli. 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. Terminator-cloning shuttle vector using the expression of chloramphenicol acetyltransferase as the reporter. 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 under control of the vegII promoter. The order of the major features in the plasmid is: To terminator - vegII promoter - HindIII/MCS/EcoRI - cat - rrnB terminator - ampR - pMB1 ori - pUB110 ori - neoR - bleR. |
