Re: [DIYbio] Do plasmids introduced via electroporation replicate with subsequent cell divisions?

There will always be a natural selection against the plasmid that may cause eventual deletion. Single cell organisms base their foundation on keeping everything very simple so adding a large amount of unnecessary DNA will take more energy to synthesize and maintain than just the needed chromosomal DNA. That being said, there are strains of bacteria (E.coli DH5-alpha is a common one) that are engineered to preserve plasmids over a long period of time.

Typically with a system like algae, plasmids will be constructed and preserved in an E.coli host strain and then moved over to the desired algae strain. I don't know how well an algae strain will preserve a plasmid over time, but if that is a concern (you need to maintain the culture at room temperature or above for a long period of time and the plasmid does not contain information necessary for survival) inserting directly to the genome with a CRISPR technique could be possible. You may be able to skip the plasmid insertion all together by conjugating CRISPR with a TEF2 like protein, but I would have to do some research to really say how.

-SG

On Tue, Sep 19, 2017 at 11:32 AM, Cory J. Geesaman <co...@geesaman.com> wrote:

What about within things like algae?

On Tuesday, September 19, 2017 at 12:49:55 PM UTC-4, Rikke wrote:

In bacteria, at least, the plasmids are replicated with the cell cycle - it's how "we" make more plasmid. =)

On Tue, Sep 19, 2017 at 9:33 AM, Cory Geesaman <co...@geesaman.com> wrote:

The title pretty much has the substance of this.  I'm curious if you can introduce plasmids to a cell via electroporation and have those plasmids replicated in subsequent cell divisions, or if they only end up in one of the cells while multi-generational versions must be incorporated into the DNA somehow.  Would the answer differ for different types of cells?

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