|
 zouden
Neuroscientist
 
Registered: 11/12/07
Posts: 7,091
Loc: Australia
Last seen: 2 years, 7 months
|
 Re: Glowing Mushrooms [Re: Zepplin]
#8491692 - 06/06/08 04:43 AM (3 years, 11 months ago) |
|
|
Wow, great photos! I had no idea bioluminescent fungi looked that good. Are these foxfire, or is that a different thing?
--------------------
I know... that just the smallest
part of the world belongs to me
You know... I'm not a blind man
but truth is the hardest thing to see
|
 Gumby
Fishnologist
Registered: 06/13/01
Posts: 23,976
Loc: BRICK CITY, NIGGA! 
Last seen: 1 minute, 49 seconds
|
Re: Glowing Mushrooms [Re: zouden]
#8492007 - 06/06/08 08:05 AM (3 years, 11 months ago) |
|
|
They're different, see my above post.
|
perosiste
Overcomplicates shit for fun
Registered: 03/26/08
Posts: 1,030
Last seen: 2 months, 20 days
|
Re: Glowing Mushrooms [Re: Zepplin]
#8492027 - 06/06/08 08:13 AM (3 years, 11 months ago) |
|
|
I wonder if the glow is species specific or if it has something to do with the substrate
--------------------
Risk Management will be the downfall of western civilization
|
 wisp
Registered: 04/13/08
Posts: 5,304
|
Re: Glowing Mushrooms [Re: perosiste]
#8494926 - 06/06/08 11:44 PM (3 years, 11 months ago) |
|
|
Species specific. If you find a substrate that makes non-bioluminescent fungi glow, you will be a rich man.
|
zouden
Neuroscientist
Registered: 11/12/07
Posts: 7,091
Loc: Australia
Last seen: 2 years, 7 months
|
Re: Glowing Mushrooms [Re: wisp]
#8495014 - 06/07/08 12:17 AM (3 years, 11 months ago) |
|
|
The gene for luciferase is easily spliced into other organisms. But it still needs the substrate luciferin, which presumably these fungi can also make. Transplanting that from one organism to another would probably be more difficult as I assume it's more than one gene.
But if you could do it, you could make any species luminescent...
--------------------
I know... that just the smallest
part of the world belongs to me
You know... I'm not a blind man
but truth is the hardest thing to see
|
wisp
Registered: 04/13/08
Posts: 5,304
|
Re: Glowing Mushrooms [Re: zouden]
#8495029 - 06/07/08 12:23 AM (3 years, 11 months ago) |
|
|
It's been done with numerous organisms already. It's used as a biomarker, but then it got out into the pet trade for novelty purposes.
But it's quite different to giving fungi a nutrient that makes them glow.
|
zouden
Neuroscientist
Registered: 11/12/07
Posts: 7,091
Loc: Australia
Last seen: 2 years, 7 months
|
Re: Glowing Mushrooms [Re: wisp]
#8495067 - 06/07/08 12:40 AM (3 years, 11 months ago) |
|
|
Really? Luciferase in pets? Do you have to feed them luciferin?
--------------------
I know... that just the smallest
part of the world belongs to me
You know... I'm not a blind man
but truth is the hardest thing to see
|
wisp
Registered: 04/13/08
Posts: 5,304
|
Re: Glowing Mushrooms [Re: zouden]
#8495098 - 06/07/08 12:59 AM (3 years, 11 months ago) |
|
|
Sorry I don't think they've used luciferin as such, but the green fluorescent protein found in jellyfish. They've used it on fish, rabbits and pigs. So I suppose technically they aren't bioluminescent but fluorescent.
Luciferin/luciferase though are used as biomarkers.
|
zouden
Neuroscientist
Registered: 11/12/07
Posts: 7,091
Loc: Australia
Last seen: 2 years, 7 months
|
Re: Glowing Mushrooms [Re: wisp]
#8495135 - 06/07/08 01:11 AM (3 years, 11 months ago) |
|
|
Yes, I've used it as a biomarker by introducing the luciferase gene, and then adding luciferin when I want to do the assay. I'm wondering if anyone has managed to transplant the luciferin gene(s) so that it doesn't have to be added. I don't think so, but it would be pretty cool.
--------------------
I know... that just the smallest
part of the world belongs to me
You know... I'm not a blind man
but truth is the hardest thing to see
|
wisp
Registered: 04/13/08
Posts: 5,304
|
Re: Glowing Mushrooms [Re: zouden]
#8495159 - 06/07/08 01:22 AM (3 years, 11 months ago) |
|
|
It would but I'm not really into transgenics without very good reason. It has to be a compelling arguement for me to see otherwise.
Did you need to introduce the gene at the 4-cell embryo stage?
|
zouden
Neuroscientist
Registered: 11/12/07
Posts: 7,091
Loc: Australia
Last seen: 2 years, 7 months
|
Re: Glowing Mushrooms [Re: wisp]
#8495293 - 06/07/08 02:31 AM (3 years, 11 months ago) |
|
|
Actually it was sugarcane cells, shot with a gene gun. The transgene wasn't inherited by daughter cells, only the cells that were hit expressed the luciferase.
If I were doing it to an animal I guess it's done at the 4-cell embryo stage? But I don't really know.
--------------------
I know... that just the smallest
part of the world belongs to me
You know... I'm not a blind man
but truth is the hardest thing to see
|
wisp
Registered: 04/13/08
Posts: 5,304
|
Re: Glowing Mushrooms [Re: zouden]
#8495400 - 06/07/08 03:53 AM (3 years, 11 months ago) |
|
|
Yes to my knowledge all animals need to be done at the 4-cell embryo stage. Obviously though, messing with an embryo when so small has enormous fatality rates. Doing it at that stage though allows the gene to be incorpated into the genome of the host.
So with plants then the transgene can be transposed at anytime? If it is not heritable does that mean then that the gene is not really incorporated into the genome of the plant? That's really quite interesting.
|
zouden
Neuroscientist
Registered: 11/12/07
Posts: 7,091
Loc: Australia
Last seen: 2 years, 7 months
|
Re: Glowing Mushrooms [Re: wisp]
#8495501 - 06/07/08 05:00 AM (3 years, 11 months ago) |
|
|
The gene can be incorporated into the genome, but it doesn't have to be for it to be expressed. So if you throw a whole lot of DNA into the nucleus of a cell, it will be expressed as long as that DNA is there (which was 3-4 days for the plant and animal cells I've worked on). It's called 'transient expression' and it's pretty easy. If you want stable expression, you just select for the cells that have the DNA (by introducing a herbicide/antibiotic resistance gene) and wait longer than the 3-4 days. Most of the cells will die, but the ones that integrated the DNA into their chromosomes will survive.
So it just depends on whether you want to study the gene or protein product, or if you actually want to create a transgenic organism. If it's the former, you'd go with transient expression because it's really easy. For plants you use a gene gun, for animal cells you use a chemical called lipofectamine.
--------------------
I know... that just the smallest
part of the world belongs to me
You know... I'm not a blind man
but truth is the hardest thing to see
|
wisp
Registered: 04/13/08
Posts: 5,304
|
Re: Glowing Mushrooms [Re: zouden]
#8495513 - 06/07/08 05:11 AM (3 years, 11 months ago) |
|
|
Wow, interesting. How the hell does that gene gun actually target a single cell, let alone the nucleus?
It is incredible what science can do these days. How does lipofectamine work?
The example I was learning about in relation to the green fluorenscent gene was to allow the development of specific muscles or organs to be followed, which is why creating a transgenic organism was necessary.
|
zouden
Neuroscientist
Registered: 11/12/07
Posts: 7,091
Loc: Australia
Last seen: 2 years, 7 months
|
Re: Glowing Mushrooms [Re: wisp]
#8495544 - 06/07/08 05:34 AM (3 years, 11 months ago) |
|
|
Yes, that makes sense, in fact it sounds familiar now that you mention it.
The gene gun doesn't target a single cell, you load it with hundreds of tungsten particles and it sprays them like a shotgun. If you do it with a gene that produces a coloured product, you can actually see all these coloured cells spread across the tissue like freckles. You can also use gold particles apparently. The DNA sticks to the metal particles and gets hurled through the membrane and, if you're lucky, into the nucleus.
Lipofectamine (and competing products like Fugene and Effectene - I prefer Effectene) is a strange type of oily chemical which temporarily permeabilises the membrane (and nuclear membrane) enabling DNA to seep in. It's expensive and highly flammable, according to the bottle.
Another way to do it is to electrocute the cells, temporarily making the membrane permeable. It's called electroporation and it works for all kinds of cells but I get the impression it's not very effective because it seems to be a method of last resort. I've only done it to bacteria.
 
The one on the right is the model I used.
--------------------
I know... that just the smallest
part of the world belongs to me
You know... I'm not a blind man
but truth is the hardest thing to see
|
wisp
Registered: 04/13/08
Posts: 5,304
|
Re: Glowing Mushrooms [Re: zouden]
#8495565 - 06/07/08 05:55 AM (3 years, 11 months ago) |
|
|
Expensive looking bits of equipment.
Right, the way that gene gun works makes more sense than targeting a single cell. Shooting bits of metal into a cell, does that cause significant cell mortality?
Also would electroporation have sifnificant cell mortalities, or is it a very small charge?
What kind of research have you used these methods in, if you don't mind me asking?
|
zouden
Neuroscientist
Registered: 11/12/07
Posts: 7,091
Loc: Australia
Last seen: 2 years, 7 months
|
Re: Glowing Mushrooms [Re: wisp]
#8495690 - 06/07/08 08:08 AM (3 years, 11 months ago) |
|
|
Both those methods have high cell mortality. Lipofectamine is much better, but less versatile. Cell mortality usually isn't an issue because there's no shortage of cells for this kind of work.
A brief list of things I've genetically engineered:
-sugarcane with gene gun: we were making a transgenic crop that produces a more complex carbohydrate than just sucrose
-bacteria with electroporation or heat-shock: various projects
-algae with glass bead vortexing: making a strain that can produce hydrogen gas through photosynthesis
-mammalian cells with lipofectamine: testing DNA before introduction into mouse - I used GFP as a marker, among other things
-mouse line with embryonic microinjection: I haven't done this personally, I'm outsourcing this to a specialist company. Roughly $2000 per mouse line.
The main methods that I haven't done are Agrobacterium-mediated gene transfer into plants and fungi, and viral vectors.
--------------------
I know... that just the smallest
part of the world belongs to me
You know... I'm not a blind man
but truth is the hardest thing to see
|
wisp
Registered: 04/13/08
Posts: 5,304
|
Re: Glowing Mushrooms [Re: zouden]
#8502763 - 06/09/08 06:07 AM (3 years, 11 months ago) |
|
|
Yes I suppose cell mortality isn't really an issue considering how small cells are and how many are even in a tiny piece of tissue.
It seems they make trangenic organisms for a massive range of applications. How do you feel about it morally/ethically?
|
zouden
Neuroscientist
Registered: 11/12/07
Posts: 7,091
Loc: Australia
Last seen: 2 years, 7 months
|
Re: Glowing Mushrooms [Re: wisp]
#8502777 - 06/09/08 06:20 AM (3 years, 11 months ago) |
|
|
I don't have a problem with it. I don't see any difference (from a moral or ethical stance) between modern genetic engineering and classical breeding. I mean, we have dog breeds that are more susceptible to back problems or liver disease, so it's not like classical breeding can't lead to reduced quality of life.
As long as the technology is used responsibly, I'm all for it. And in the areas I've worked, it's always used responsibly (and there's very heavy regulation on it in Australia) - but I can't speak for work going on in corporations. Science and technology is neutral, but people aren't necessarily so - particularly when profits are on the line.
--------------------
I know... that just the smallest
part of the world belongs to me
You know... I'm not a blind man
but truth is the hardest thing to see
|
wisp
Registered: 04/13/08
Posts: 5,304
|
Re: Glowing Mushrooms [Re: zouden]
#8502808 - 06/09/08 06:40 AM (3 years, 11 months ago) |
|
|
It's not so much that than the unknown side-effects of modifying organisms. No-one can predict what will happen. GM crops are an example of problems arising from GMO's. But more problematic than the social issues that arise from it are the potential biological issues of introducing modified DNA into wild populations. Selectively breeding dogs is still breeding within the one species. Modifying an organism to express trait from an entirely different phyla or kingdom is very different indeed.
|
|
Previous
Index
Next
Threaded
Edit 
Reply 
Anonymous
