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. 2011 Jun 30;7(6):e1002118. doi: 10.1371/journal.ppat.1002118

Table 3. Plasmids used in this study.

Plasmid Description Reference or source
pMM47.A ori ColE1, ori pCC7, Apr ,Cfr, E. coli - C. canimorsus expression shuttle vector. [33]
pPM1 pMM47.A where the ermF promoter has been replaced by the stronger gpd promoter: 117 bp upstream of the gpdC ORF start codon were amplified with primers 5081 and 5469 and cloned into pMM47.A using SalI and NcoI restriction sites. This study
pPM2 Full length gpdC containing its putative promoter region amplified with primers 5081 and 5082 and cloned into pMM47.A using SalI and SpeI restriction sites. This study
pPM3 Full length gpdC with a C-terminal His-Strep double tag amplified by 2-step overlapping PCR with primers 5081, 5467 and 5530 and cloned into pMM47.A using SalI and SpeI restriction sites. This study
pFR4 Full length gpdD amplified with primers 6133 and 6057 and cloned in pPM1 using NcoI and XbaI restriction sites. This study
pFR5 Full length gpdG amplified with primers 5008 and 6055 and cloned in pPM1 using NcoI and XbaI restriction sites. This study
pFR6 Full length gpdE amplified with primers 5959 and 5060 and cloned in pPM1 using NcoI and XbaI restriction sites. This study
pFR7 Full length gpdF amplified with primers 5062 and 5063 and cloned in pPM1 using NcoI and XbaI restriction sites. This study
pFR8 Full length gpdD with a C17G point mutation amplified with primers 6056 and 6057 and cloned in pPM1 using NcoI and XbaI restriction sites. This study
pFR9 Full length gpdG with a C21G point mutation amplified with primers 6054 and 6055 and cloned in pPM1 using NcoI and XbaI restriction sites. This study
pFR10 Full length gpdG with a E205G point mutation amplified by overlapping PCR using primers 5008/6061 and 6060/6055 and cloned in pPM1 using NcoI and XbaI restriction sites. This study
pMM121.1 Full length siaC amplified by inverse PCR using primers 5045+5046 on pMM52 as a template to insert to C17Y substitution in siaC. This study
pMM25 ori ColE1, Kmr, Cfr.Suicide vector for C. canimorsus. [33]
pMM52 Full length siaC with a C-terminal His tag cloned in pMM47.A using NcoI and XbaI restriction sites. [13]
pMM106 ori ColE1 , Kmr , Cfr , EryR , Mutator plasmid for the replacement of siaC by ermF [33]