We have a strain of Lactococcus lactis transformed with a plasmid to express an heterologous protein. We are planning to use this strain to express another protein using the same vector, which features a constitutive promoter. Is it feasible? Can be "re-electroporated"?
Thanks in advance!
You can "re-electroporate" an already transformed strain, but you should use a different vector with a different antibiotic marker. If you use the same vector with the same antibiotic marker you won't be able to select for your new transformants.
I didn't realized that. Fortunately, I can choose between two different antibiotic markers, thank you!
Eoghan provided a good answer. I just would like to add:
- because of the efficiency of electroporation, it is also feasible to directly co-transform an electrocompetent strain with two plasmids (plate must then contain both antibiotics).
- if possible, it is preferable to use vectors with different origins of replication to maintain both plasmids
Could you clone the gene encoding your new protein of interest into the existing plasmid therefore expressing both proteins from a single plasmid? Seems more efficient. Also, using different plasmids with different origins of replication (which as Cedric has correctly mentioned you will need to do) may result in different protein levels due to differences in plasmid copy number rather than differences in promoters - depends whether this is relevant or not for your particular experiment.
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