Restriction digests of the clone give the following sizes (kb): PstI--20.0; HindIII--20.0; BglII--20.0; EcoRI--17.0; 2.6. This vector transfers efficiently to a broad host range including Escherichia coli HB101, Methylobacterium organophilum DSM 761, Pseudomonas AM1, Pseudomonas putida, and Pseudomonas aeruginosa. IMPORTANT: To prevent amplification of a rearranged and/or deleted cosmid, we recommend streaking on LB + amp plates at 30C and picking small colonies for liquid culture. This is a cosmid with a broad host range that can be used to characterize the genome of C1-utilizing microorganisms. This was constructed from pLA2917 (ATCC 39840) by deleting the kanamycin resistance segment from Tn5 to avoid internal rearrangement when the transposon moves and to permit selection of conjugants that have a Tn5 insertion. |
