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. 2007 Oct 24;2(10):e1055. doi: 10.1371/journal.pone.0001055

Figure 1. Experimental design to measure recombination rate at different positions of the R. solanacearum chromosome.

Figure 1

Target positions on the chromosome (TCP) and on the megaplasmid (TMP) were identified and amplified by PCR. PCR products from the different genomic positions were cloned in appropriate vectors and afterwards, labeled with the aacC3-IV gene (gentamycin cassette). The pTCP (versus pTMP) plasmid carrying homologous GMI1000 fragments were linearized and resulting plasmids were used as donor to transform naturally the wild type strain GM1000 and recombination rate of each position designed. Total genomic DNA from R. solanacearum transformants and carrying aacC3-IV cassette resulting from double crossing-over were used as exogenous DNA donor to “re-transform” the wild type strain GM1000 and the CFBP2968, NCPPB332 and CFBP2957 strains to determine the recombination rate of genomic DNA. (Amp, ampicillin and Kn, kanamycin).