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. 2018 Jun 26;9:1329. doi: 10.3389/fmicb.2018.01329

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Copyright © 2018 Beker, Rose, Lykkebo and Douthwaite.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Identification of ICE-related Pasteurellaceae genes using multiplex PCR. The gels show the analyses of 75 field isolates, 43 of which were P. multocida (identified by the 600 bp band that is specific for Pasteurella spp.) and 32 were M. haemolytica (band at 720 bp and lack of 600 bp band). The four ICE-related PCR products correspond to parB (503 bp), the ICE-specific relaxase gene (437 bp), and the longer and shorter versions of the integrase genes int1 (301 bp) and int2 (215 bp). In strains that lacked all these ICE genes (e.g., Pmu13083) a faint artifact band of 250 bp was sometimes apparent. This band was no longer evident after raising the primer annealing temperature 3°C above the optimal hybridization temperature for the canonical sites. The control lane (-DNA) shows a reaction without template DNA. The bands in the GeneRuler ladder on the left are in 100-bp steps.