Many numbers for E. coli: https://bionumbers.hms.harvard.edu/search.aspx?trm=coli
Everything You Ever Wanted to Know about E. Coli (2008)
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Re: Everything You Ever Wanted to Know about E. Coli (2008)
#12I use E.coli on a pretty daily basis for cloning. It's alright, and there is so much work that has gone into it as a chassis organism, but overall there are definitely better organisms out there if we just took the time to figure them out (Vibrio natriegens and Bacillus subtilis are two examples). We absolutely do not have a clear idea of how E.coli works. Hell, we don't even know how almost 1/3 of the genes work on…
Ah man. Back in my university days, I worked in an infectious disease (HIV) lab that shared space with some GI docs. Went down to Mexico and came back with enteropathogenic e. coli. While they were super happy to get some fun samples and blood works, I've never felt sicker in my life.
My side would immortalize b-cells with Epstein-Barr... but that experience left me with a healthy respect for e coli.
Re: Everything You Ever Wanted to Know about E. Coli (2008)
#13Earlier quoted context omitted.
Normal E.coli doubling time, which means about half the speed of Vibrio natriegens. I use NEB turbos sometimes, but they still don't bring down the liquid culture time down from overnight to within a day, mostly. I would want E.coli to sporulate because then shipping and distribution of strains is a lot easier. This was a problem back when I was shipping at FreeGenes and still is a problem. I used Bacillus subtilis t…
If all you're interested is cloning/expression, then I guess that makes sense, although I've not heard anyone complain that 20 min is a slow doubling time. can't you just inoculate your starting culture with a larger amount or something? I often find myself starting cultures late in the day so that they don't overgrow the next day (although I am not doing expression work). Vibrio natriegens seems great for your purpo…
> can't you just inoculate your starting culture with a larger amount or something
No, I'm doing high throughput cloning in such a way where a limiting factor is going from 1 cell to X cells (usually pickable colonies). So liquid culture doesn't matter quite as much, since I can sequence validate from a colony split (half into colony PCR, half into new culture), and sequencing takes approximately as much time as the growth step takes.
> I'm curious if there is a tradeoff to its super fast growth. Does it have a greater mutation rate?
Eh, not really. The biggest tradeoff is that it isn't studied nearly as much as E.coli, so the chemical competence protocols kind of suck. I can't actually use it in my pipeline right now because of that. So I'm stuck with E.coli until I can figure out a good transformation protocol.