Characterization of chimeric plasmid cloning vehicles in Bacillus subtilis

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Abstract
Restriction endonuclease cleavage maps of 7 chimeric plasmids that may be used for molecular cloning in B. subtilis are presented. These plasmids all carry multiple antibiotic resistance markers and were constructed by in vitro molecular cloning techniques. Several of the antibiotic resistance markers were shown to undergo insertional inactivation at specific restriction endonuclease sites. Kanamycin inactivation occurred at the BglII site of pUB110 derivatives, erythromycin inactivation occurred at the HpaI and BclI sites of pE194 derivatives and streptomycin inactivation occurred at the HindIII site of pSA0501 derivatives. A stable mini-derivative of pBD12 was isolated and characterized. By using these plasmids, proteins involved in plasmid-coded kanamycin and erythromycin resistance were identified. The properties and uses of these chimeric plasmids in the further development of recombinant DNA technology in B. subtilis are discussed.