Membrane topology of conserved components of the

Membrane topology of conserved components of the type III secretion system from the plant pathogen Xanthomonas campestris pv. vesicatoria By: Carolin Berger, Guillaume P. Robin, Ulla Bonas and Ralf Koebnik Supplementary Table S1. Oligonucleotides used in this study

Restriction sites are underlined. Start and stop codons are highlighted in green and pink, respectively. Name

Sequence (5′–3′)

PCR amplification and cloning of hrc genes into pCR 2.1-TOPO CCG GGA ATT CAC CAT GAC CAT GCA GCT TCG CGT AAT Topo-HrcD-5′ GAC CCC AAG CTT ATT GCG CCG CTT GCT GCG GCA GGG ATG Topo-HrcD-3′ TC CCG GGA ATT CAC CAT GCA GAT GCC TGA CGT CGG CTC Topo-HrcR-5′ G CCC AAG CTT ACC GAT AGC TCA GAA CCA GGC CGT G Topo-HrcR-3′ CCG GGA ATT CAC CAT GGA CCA TGA CGA TCT AGT Topo-HrcS-5′ GCG CCC AAG CTT ATG GGA ACG CCG CCT GCA TCA ATG C Topo-HrcS-3′ CCG GGA ATT CAC CAT GAA CGA CGC CAC CGA AGC Topo-HrcT-5′ CTT GC CCC AAG CTT AGT GCG TCG CGC CTC CCT TGC CAG ACA Topo-HrcT-3′ G CTA GCT AGC AGG AGG AAT TCA CCA TGT CGG AAG Topo-HrcU-5′ AAA AAA CCG AGA AGC CG CCC AAG CTT AGC ATG GCA GGG CTC CAT CGC CGT CGC Topo-HrcU-3′ CGT GGT C CCG GGA ATT CAC CAT GCG AGC GAC AAG GTA TTT Topo-HrcV-5′ CGC CCC AAG CTT ACA CCA CCA CTC TAC CGA TCG GCT GCA Topo-HrcV-3′ GCT G Mapping of Tnpholac1 insertions (colony PCR and DNA sequencing) Seq-Tnpholac1 CGC CGG GTG CAG TAA TAT CG CTG TTT CTC CAT ACC CGT T pBAD-5′ Construction of site-directed reporter fusions (cloning into pDT) CTA GCT AGC AGG AGG AAT TCA CCA TGA CCA TGC Topo-HrcD-NheI-5′ AGC TTC GCG TAA TG GCT CTA GAG CAT AGC ACC AGC GGG CCG AAG CG Topo-HrcD-C80-3′ GCT CTA GAG CCG GAA ACC GAG AAA ACG CC Topo-HrcD-G239-3′ CTA GCT AGC AGG AGG AAT TCA CCA TGG ACC ATG Topo-HrcS-NheI-5′ ACG ATC TAG TGC G GCT CTA GAC GAG ATC GAT GCA TCC TGC Topo-HrcS-S50-3′ GCT CTA GAC AAT GCC TGG CCG AAC TGC Topo-HrcS-L80-3′ GCT CTA GAT GGG AAC GCC GCC TGC ATC AAT GC Topo-HrcS-P86-3′ CTA GCT AGC AGG AGG AAT TCA CCA TGA ACG ACG Topo-HrcT-NheI-5′ CCA CCG AAG CCT TGC Topo-HrcT- Q119-3′ GCT CTA GAC TGC ACA TTG TTG TAA CCG GC GCT CTA GAC GAC GGT TCG AGT TTA TCG GC Topo-HrcT- S223-3′ GCT CTA GAG ATT GGC TGA CTC AAG TTC GAC G Topo-HrcT- I229-3′ CTA GCT AGC AGG AGG AAT TCA CCA TGT CGG AAG Topo-HrcU-5′ AAA AAA CCG AGA AGC CG GCT CTA GAC GAA AAG AGT TTC TTG ACC CCG GTT GC Topo-HrcU-S134-3′

Restriction sites EcoRI HindIII EcoRI HindIII EcoRI HindIII EcoRI HindIII NheI, EcoRI HindIII EcoRI HindIII

NheI, EcoRI XbaI XbaI NheI, EcoRI XbaI XbaI XbaI NheI, EcoRI XbaI XbaI XbaI NheI, EcoRI XbaI