was: glowing plant - Open Source chloroplast engineering

[Andreas "Mega" Stuermer]

Hi guys, 

I am currently re-visiting the idea of chloroplast engineering. Chloroplast engineering yields loads of proteins, and is only maternally inherited (containment bonus). Antibiotic selection marker will be removable with Cre-Lox, and should happen with low frequency by iself too. 

First project: creating the empty vector, SciHouse in Florida will store the vector in bulk and share with anyone interested (open source) 

Second project: putting the lux operon in, just because. https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0015461 *

I have seen one of these plants, and they were visibly glowing to the naked eye (but weak). Note that that was way before the Silicon Valley glowing plant cowboy project. 

*Unfortunately they stopped selling them in the United States for no apparent reason (maybe they didnt't get enough customers).  

Not sure if I should start with weeping willow or tobacco. I've been routinely working with tobacco and can share my protocols with our US collaborators so they can do the tissue culture there. Tobacco is pretty lame though. 

Anyways, just wanted to see if anyone would be willing to chime in, synthesis is expected to cost around 500$. Attached the file. Addition of lux will just be one PCR step 

[Nathan McCorkle]

I have seed from my gleaux plant, a bunch that dropped into the pot
that it was planted in have sprouted so I know they're viable. I never
saw the original glow, nor these... and they don't show up with a 30
second dSLR exposure image. I had been working to setup my laptop to
remotely trigger my camera to take a series of images and then add a
bunch of images together to try stack the brightness... but I got
caught up with other things and then my laptop went through 3 weeks of
failed ubuntu upgrade attempts before I got it straightened out again
recently.

Anyway, if you think you could use some seed, how many would you want?

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-Nathan

[Andreas Stuermer]

Now that's interesting, I swear I saw it glow. It was the first generation (bioglow) before they called it gleaux. 

If you can send it to my friend in Florida, that would be cool. He might try to grow it on spectinomycin, which was their selection marker. Maybe they didn't delete it

I will check if we can grow it in the museum's biolab or if that requires months of approval. We might be lucky because we already had approval to put gfp and lux into tobacco. 

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[John Griessen]

On 10/26/18 7:57 PM, Andreas "Mega" Stuermer wrote:
> First project: creating the empty vector, SciHouse in Florida will store the vector in bulk and share with anyone interested (open
> source)

How do you do transformation? Is electroporation used? I'd like more beta testers of my electroporator.
So far, the folks that have gotten one have never used it on live cells. Kind of slows the project momentum...

[Andreas Stuermer]

Gene gun has the advantage that you can shoot it into leaf tissue and then regenerate it. 

Electroporation of protoplasts may be possible but they are easy to burst

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