[koeng]

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 to make the sporenet protocol which worked, but the vector backbone switching was pretty painful.

Clinical strains == not necessarily GRAS. Also, they're clinical, so regulated by MTAs, which are a big no-no.

I could make the E11K purB mutation, but why would I go through the effort in an inferior species of bacteria when I can invest my time into something like Vibrio natriegens? The payoff is better at the end.

[pazimzadeh]

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 purposes. I'm curious if there is a tradeoff to its super fast growth. Does it have a greater mutation rate?

[koeng]

> 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.