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. 2002 Jan;68(1):306–315. doi: 10.1128/AEM.68.1.306-315.2002

FIG. 8.

FIG. 8.

PCR analysis of site of integration in pBNG3.0 transformants. (A) Restriction endonuclease map of pBNG3.0. Only restriction endonuclease sites relevant to this study are shown. Template DNA was prepared using a rapid boiling procedure. Ten GUS+ transformants (lanes 1 to 6 and 8 to 11) and one GUS− transformant (lane 7) were analyzed for both 5′ and 3′ nor-1 integration. (B) Schematic for 5′ nor-1 integration with PCR data. 5′ nor-1 integrants resulted in a 3.1-kb PCR fragment (lanes 1, 2, 5, 8, 11) with PCR1 primers. (C) Schematic for 3′ nor-1 integration with PCR data. 3′ nor-1 integrants resulted in a 2.1-kb PCR fragment (lanes 3, 4, 6, 9, 10) with PCR2 primers. All GUS+ transformants were 5′ or 3′ nor-1 integrants while the GUS− transformant (lane 7) was negative for both 5′ (9B) and 3′ (9C) nor-1 integration. Lanes labeled M in panels B and C represent molecular size markers.