The cloning of the 5.4-KB plasmid of Bacillus megaterium
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The construction of a stable cloning vector would be very useful to the study of genetics of Bacillus megaterium. It seems reasonable to assume that a vector made with a natural resident plasmid of this species will be stable. This is the rationale behind cloning the 5.4-kilobase plasmid of B. megaterium. The 5.4-kb plasmid and the vector were each cut with a restriction endonuclease that produced blunt ends. These two plasmids were ligated using the enzyme T4 DNA ligase. The recombinant plasmid was transformed into E. coli. Transformant DNA was then analyzed using restriction enzvmes. Because of errors in the restriction analysis data used, only part of the 5.4-kb plasmid has been cloned. However, this hybrid plasmid may still be useful in constructing cloning vectors for use in B. megaterium. and other bacilli.