Team:Emory/Experiments
EXPERIMENTS
Here, we've listed the experiments we conducted throughout this investigation. To the right is a graphic which describes the overall process we followed for experimentation. Each individual experiment corresponds to a certain section of the graphic.
Experiment iG1: The purpose of this experiment was to subclone pWH1266 plasmid origin into pSB1C3. The pWH1266 plasmid origin is specifically designed for Acinetobacter plasmids.
Experiment iG2: The purpose of this experiment was to create the shuttle vector pWH1266 ori-Ptac-lacOc-R67-DHFR-pSB1C3 by subcloning the Ptac-lacOc-R67-DHFR insert into the pWH1266 ori-pSB1C3. This shuttle vector has trimethoprim resistance.
Experiment iG3: The purpose of this experiment was to create the shuttle pWH1266 ori-Ptac-lacOc-aph(3')-VIa-pSB1C3 by subcloning the Ptac-lacOc-aph(3')-VIa insert into the pWH1266 ori-pSB1C3.
Experiment iG4: The purpose of this experiment was to create the shuttle vector pWH1266 ori-Ptac-lacOc-ampC ADC-33-pSB1C3 by subcloning Ptac-lacOc-ampC ADC-33 insert into the pWH1266 ori-pSB1C3. This shuttle vector has ceftazidime resistance.
Experiment iG5: The purpose of this experiment was to create the shuttle vector pWH1266 ori-tetR-PtetA-tetM-pSB1C3 by subcloning the tetR-PtetA-tetM insert into the pWH1266 ori-pSB1C3.
Experiment iG6: The purpose of this experiment was to insert the gusA reporter gene, lacI-PT5-gusA, into pWH1266 ori-Ptac-lacOc-ampC ADC-33-pSB1C3 to create pWH1266 ori-Ptac-lacOc-ampC ADC-33-lacI-PT5-gusA-pSB1C3.
Experiment iG7: The purpose of this experiment was to insert the gusA reporter gene, lacI-PT5-gusA, into shuttle pWH1266 ori-Ptac-lacOc-aph(3')-VIa-pSB1C3 to shuttle pWH1266 ori-Ptac-lacOc-aph(3')-VIa- lacI-PT5-gusA-pSB1C3
Experiment iG8: The purpose of this experiment was to repeat experiment iG6 to ensure the validity of its results.
Experiment iG9: The purpose of this experiment was to show that we could do a plasmid preparation from A. baylyi and use it to transform E. coli and A. baylyi. This supports the use of A. baylyi as an alternative vehicle for transformation.
Experiment iG10: The purpose of this experiment was to subclone the mambalgin into our shuttle vector pWH1266 ori-Ptac-lacOc-ampC ADC-33-pSB1C3.
