I am trying to figure out how temperature sensitive plasmids are made/isolated and what the biochemical/cellular mechanism is that causes them to be lost/lose function at higher temperatures.
https://www.researchgate.net/post/why_plasmid_from_Agrobacterium_lost_at_higher_temperatures
This is done to easily get rid of the plasmid after you are done with recombineering (otherwise it will be a bit of a hassle to grow the cells non-selectively then screen for plasmid loss). The basis of the temp sensitivity is a temperature sensitive replication protein (rep) carried on the plasmid and required for its replication, at non-permissive temps this becomes nonfunctional forcing the loss of the plasmid from the cells.
The mentioned plasmid contains a temperature-sensitive origin of replication.
Kindly check these links for a database and a research article
http://parts.igem.org/Part:BBa_K1223012
https://www.pnas.org/content/97/12/6640
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