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. 2005 Jun;71(6):3068–3076. doi: 10.1128/AEM.71.6.3068-3076.2005

FIG. 2.

FIG. 2.

Identification of pBM300 as the source plasmid for clone III by Southern hybridization and separation of pBM300 into PV361. (A) Agarose gel electrophoresis of plasmids. QM B1551, the wild-type strain, contains all seven plasmids; QM B1551/26::spc contains plasmids isolated after spc gene insertion; the PV203 strain does not contain plasmid pBM300 (used as a negative control); and PV634 contains only pBM300::spc. The DNA was transferred to a nitrocellulose membrane. Then the membrane was hybridized with different probes, as indicated above panels B to E, sequentially after the old probe was stripped. (B to E). Hybridization with plasmids and derivatives from B. megaterium QM B1551 with the following specific probes: pBM500 (clone I) (40) (B), pBM400 (clone II) (38) (C), a PCR fragment specific for the clone III probe (distal to the rep gene in pBM300) (D), and the spc gene probe (E). Note that the blots in panels B to E were derived from the gel shown in panel A.