I need to select an appropriate plasmid to transform several different plant species. Ideally, I would like to use the same vector for all the species to limit variable differences and time. What is the best way to go about finding existing vectors that will work across multiple plant species while also filtering for specific elements I would like to use (e.g. nptII resistance and GFP/GUS markers)?
I would appreciate advice on common mistakes and important aspects to consider while I search, but I am mainly interested in efficient and reliable steps to follow.
Thanks in advance!
Use a T7promoter system. That way as long as you get some expression of T7 RNA polymerase-NLS, you can then drive the transcription factor using a tandem promoter to get more consistent expression. You can then drive your genes of interest using T7 promoters of defined strengths.
Using KanR for G418 selection is also an option.
You may also want to use a geminivirus derived backbone so that plasmid maintenance relies less on host replication machinery for initiation.
It would take time but you can look in the literature to see what vectors have been used in your species of interest. If they are uncommon to transform this won't take too long. If they are common species, there are probably commercially available vectors such as pCAMBIA or similar. The challenge here might be finding a selective agent that works well for all your species and cultivars.
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