I would like to modify a foliar biocontrol agent so that it has an added beneficial effect, but it requires recombinant plasmid incorporation. I can't test this agent in the field if the selection system is antibiotic-based. If anyone has a recommendation how to keep this plasmid within the bacterium in an environmentally safe way, that'd be great! Thanks!
That would be the idea if I can figure out how to keep the plasmid in.
There are many options you can use, depending upon what host strain you wish to use. You could use a host strain carrying an auxotrophic marker and use the complementing gene cloned on your plasmid as the selection. This could be for an amino acid, or D-alanine, or many other similar markers. There were also some plasmids designed many years ago carrying amber suppressor tRNA and use a host strain with amber nonsense mutation in an essential host gene.
in the first step you should determine the strain.
then .
1- using Auxotrophic marker .
2-or using complementing gene cloned .
or as the method mentioned by (Michael J.)
Article Applicability of non‐antibiotic resistance marker genes in e...
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