FIG. 1.

phaD gene replacement by homologous recombination in P. oleovorans GPo1000. The upper three lines show the construction of the plasmid pUT-phaD::tet, which was used for gene replacement of phaD. A double crossover between pUT-phaD::tet and homologous DNA in the chromosome of P. oleovorans GPo1000 resulted in the phaD::tet-integrated strain GPo1001. Gray box, tet resistance gene; black bar, phaD gene; open arrows, other genes up- and downstream of the phaD gene. Abbreviations: A; Asp718, C; ClaI, E; EcoRI, N; NruI, S; StuI, Sm; SmaI; X; XmnI.