FIG. 1.
Schematic representation of the 2.143-kb rpoE::Kmr cassette construction. (a) Generation of PCR products A, B, and C from pEH8 and pBSL180. (b) Illustration of the sequential ligation of PCR products A and C and of AC and B yielding ACB, the 2.143-kb rpoE::Kmr cassette. (c) Cloning of the 2.143-kb rpoE::Kmr cassette (PCR product ACB) into the HincII site of pGEM-3Z generating pEH13. (d) Generation of pEH14 through excision of the 2.143-kb rpoE::Kmr cassette (PCR product ACB) from pEH13 with XbaI and SphI and insertion into the XbaI-SphI sites of pGP704. This plasmid was introduced into V. angustum strain S141 by conjugation for allelic exchange to produce rpoE mutant strain EH1.