FIG 7.

Strategies for plasmid insertion mutagenesis and assessment of the transcription of downstream genes used in this work. (A) Integration of pCR-BluntII-TOPO plasmid into a target gene. The hatched boxes correspond to the internal segment of the target gene. The vector is integrated into the targeted ORF by a single-crossover event. The direction of transcription from the T7 promoter (PT7) is indicated by the small thin arrows. The small black arrow pair represents the oligonucleotides used to assess the transcription of downstream genes. (B) RT-PCRs were carried out in the presence of cDNA, total RNA, and DNA using the indicated combinations of oligonucleotides amplifying overlapping regions between the T7 promoter of the pCR-BluntII-TOPO plasmid and the specified downstream genes of the indicated A. baumannii ATCC 17978 mutant derivatives. HinfI-digested Φ DNA was used as a molecular size marker (lanes M). The lengths of the products amplified by the indicated oligonucleotide pairs are reported with respect to some of the marker bands.