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pMM47.A where the ermF promoter has been replaced by the stronger gpd promoter: 117 bp upstream of the gpdC ORF start codon were amplified with primers 5081 and 5469 and cloned into pMM47.A using SalI and NcoI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]
|