Re: [DIYbio] Creating Luminous Plants - Amateur

[Dakota]

First, yes that video is fake, some good ol' fashion troll science.  In fact, someone on this group is studying that plant right now.  The plants do exist, and do respond to touch, but through another mechanism not related to ingesting lots of protein powder and hitting the gym 8 days a week.

I never have an answer for what books to read, but someone else might, and I know the same question has been posed many times, so do some back searches of old threads and just sift through them.  A college level cell biology text book is a decent place to start.

For equipment, with careful purchases you could make a decent lab with $1000 but it's important to know what to look for, and then what you are going to use it for.  Equipment honestly isn't that expensive I've found, it's reagents and consumables that add up more quickly, and have to be replaced eventually.  I'll try to work on a blog post to answer some of your questions because it's one that is asked often.

For starters, try the old DNA extraction from strawberry's or any other thing you have lying around in your fruit or vegetable bowl with salt soap n alcohol.  You learn how to lyse a cell, how to salt out DNA, then how to precipitate it.  

https://www.youtube.com/watch?v=hOpu4iN5Bh4

[Mega]

Hi,

Yet a night-time only sollution seems more achievable

Why that? AFAIK we don't really know plant promoters that switch on when dark. So "always on" promoters (constitutively) would be the choice.


You know the difference between eukaryotic gene expression and prokaryotic one? Prokaryotes (Bacteria) can read entire operons (Promoter-GeneA-GeneB-GeneC-Terminator), while eukaryotes always need one promoter before each gene.

The only surely known light pathway is the lux operon from marine bacterie. It has also already been inserted into chloroplasts (they are ~bacterial endosymbionts in plant cells, so they can express operons)


You could get pVIB plasmid and insert it into E.Coli to get quick, yet very impressing, results.


Hope that helped a bit.


Also, there will be a geneome compiler kickstarter campaign which will synthesize many designs of glowing genes for plants then and try out which one works best. 
 

[Mega]

Might I ask, where are you located?



Another addition:

For all light emitting reactions you basically need luciferin and luciferase which react to produce light.

You saw the thread about the pre-coelenterazine pepdide? This is a luciferin, and Renilla luciferase (which is broadly available) is the luciferase. So you could build a full light-generating pathway using just these two genes (adding two strong promoters). For comparison, the vbacterial lux operon needs LuxA+B (Luciferase) and Lux C+D+E+G, so in total 6 genes.

[Mega]

You surely can get the knowledge needed from the internet.
I'm studying biology and environment engineering, and if I were a regular student, I wouldn't have transformed one E.Coli yet.

Of course, in class you learn DNA replication in detail, but you can certainly suck in enough knowledge from the internet, to do some amazing experiments at home (if legal), in a DIY-community lab, hackerspace, university-lab, ....





  



On Monday, December 24, 2012 6:59:01 AM UTC+1, TJ Reece wrote:

Before I even try to make an attempt to have the slightest clue that I know what i'm talking about, or that I belong in the forum, I don't.  While I've always been fascinated by science, my education in the field of bio is limited to a level 100 Biology course.  However, since the movie "Avatar" came out I've had an interest in flourescent plants.  Every now and then I'll refer back to google revising my search to look for bioluminous plants or any modifications that are currently on the market to apply your own.  While Ideally I would like my garden plants to emit light constantly, or at a pulse like a firefly; Yet a night-time only sollution seems more achievable.  I would love to see plants or grasses in my garden illuminate at night.  The simplest sollution would be to paint the plants with UV ink, but I don't see this option to be practical. 

 

When I look into the field of genetic engineering WAY too much graduate level information comes up that I can't decipher.  I have no problem reading a book or two to understand the concepts involved in a project like this, but I prefer more trial and error methods. 

 

I did find something that looks like it might be worth a try, but I would like to get a second opinion first.  I saw this link human-plant hybrid of a youtube video that uses human hair and fern seeds to make the fern react to touch.  It just seems that this video may be fake. 

 

After looking through various organisms that have bioluminous properties, I was wondering about dinoflagellates http://en.wikipedia.org/wiki/Dinoflagellate and whether it's possible using this method above to incorporate luminous properties into something like a fern seed.  Seems a little way out of my education level to contemplate. 

 

If ANYONE on this board could give me any useful advise it would be much oblidged.  Im unsure on what terms to search for, what books would be helpful, what equipment I would need.  I could spend up to 1k, or maybe 2k on equipment AT MOST, but other than that I can't afford to finance the project.  I've been looking at DNA synthesizers, Gene Guns, and other really expensive equipment that seems a little more over the top than what I may need, but still pretty cool stuff.  Maybe I could start with plants and work my way up to mutating my own team of X-Men!!!MUAHAHAH (sorry, lol)

 

 

 

[TJ Reece]

Mega,
Did you ever make a glowing bonsai tree?  I would love to see pictures if you got them! 

[Mega]

Ok, now I seem to be even more lost than before.  I found this site to help me gain some understanding (education-portal.com) which has video classes of bio and genetics.  Suppose I wanted to specifically make grasses (festuca boulder blue) and to use the dinoflagellate luciferase gene (lux operon?).  Now, would I have to synthesize the dinoflagellates still, or could I just purchase luciferin and renila luciferase?

You would be right so far. You would extract the entire Dinoflagellate DNA and use PCR to get the lux genes needed. (You know PCR? it amplifies a piece of DNA, but not the entire DNA).
But, to get the Lux genes from PCR, you need to know the sequence to make primers (think of it like this: The entire DNA is a book, Primers are bookmarks. What is between the bookmarks is copied many many times)

Unfortunately, we don't know the lux genes form dinoglagellates. So there's no chance with this, until scientists find it out.



What already has been done: Make a PCR of  P.Phosphoreum (a glowing marine bacterium) which cut out the Lux Genes (they are very next to each other, so one PCR using 2 primers was enough), add a chloroplast-promoter and insert it into chloroplasts. They are able to express operons, and there we go. It was glowing (weakly, but visible) in fact.

See http://www.plosone.org/article/info:doi/10.1371/journal.pone.0015461

[Mega]

Did you ever make a glowing bonsai tree?  I would love to see pictures if you got them!

Unfortunately, not. Quite difficult, wanted to get a chloroplast integration plasmid once, but no lab wanted to provide me with it. That would have been quite easy, PCR -> lux genes, kanamycin resistance, -> into chloroplasts and they glow.

You can get one synthesized though. Costs ~0,35$ per base pair, makes for the chloroplast integration backbone ~ 1000 $....


You could also do 6 PCRs from ususal chloroplast DNA to make the vector, and ligate them together. That would be cheaper, but more difficult though.