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. 2018 Apr 3;9:545. doi: 10.3389/fmicb.2018.00545

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Copyright © 2018 Roy, Takamatsu, Okura, Goyette-Desjardins, Van Calsteren, Dumesnil, Gottschalk and Segura.

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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Schematic representation of mutagenesis procedures to obtain sialyltransferase substitution mutants in (A) S. suis and (B) Group B Streptococcus (GBS). (A; top part) Non-lethal mutation in neuC (Δsynth) was used first to abolish capsular polysaccharide (CPS) production. Gene substitution plasmids p4sia2,3_2 and p4sia2,3_14 were introduced into S. suis SS2Δsynth (serotype 2) and SS14Δsynth (serotype 14) mutants, respectively. (A; bottom part) In order to reintroduce a functional neuC gene into SS2Δneu2C/cps3K and SS14Δneu14C/cps3K mutants, the p4neuC plasmid was introduced into SS2Δneu2C/cps3K and SS14Δneu14C/cps3K mutants to obtain indirect substitution mutants SS2sia2,3 and SS14sia2,3. (B) In order to substitute GBS type III and V sialyltransferases, direct gene replacement by double-crossover homologous recombination system was used by introducing mutation plasmids p4sia2,6_III and p4sia2,6_V.