Evolution in quantum leaps: multiple combinatorial transfers of HPI and other genetic modules in Enterobacteriaceae - PubMed (original) (raw)

Figure 2. Sequencing approach for the intB gene of all HPI-positive isolates used in this study.

A) Region bp 41,120-45,349 of FN297818 (Box 2 in Figure 1). Amplification primers (black): pre-GI1-F and YbtS-R. Sequence primers red and black for the obtained PCR product: pre-GI1-F, IntBseq2F, IntB-F, YbtS-R, IntB-R, and IntBseq1R. PCR-positive and sequenced isolates: EHOS isolates: 01-083, 01-234, 02-195, 02-203, 02-477, 03-375, 03-525, 03-577, 04-640, R1568, 05-545. Non-EHOS isolates but with the same content in the integration hotspot: 03-273, 05-349. A non-EHOS isolate with another integration hotspot: 05-316. EHOS isolates 05-761, 06-339, H9 were PCR-positive but not sequenced because ten other EHOS isolates were sequenced and did not show polymorphisms. For the other color codes see Figure 1. B) Region bp 383–4423 of AF091251. Amplification primers (black): pre-IntB-F and YbtS-R. Sequence primers red and black for the obtained PCR product: pre-IntB-F, IntBseq2F, IntB-F, YbtS-R, IntB-R, and IntBseq1R. PCR-positive and sequenced isolates: 01-084, 01-306, 02-023, 02-272, 03-018, 03-339, 03-426, 03-595, 03-613, 03-635, 03-642, 03-739, 03-895, 05-189, 05-202, 05-680, 06-316, 10A275, 10E013, 14A001, 18D099, and X2327. From the isolates 03-093, 03-192, 03-414, and R0332 the amplification product was approximately 350 bp smaller than expected. Sequence results indicated that these isolates contained a truncated intB gene. Moreover, bp 124–470 was deleted compared to the wild-type intB gene.