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 Working with multicellular Eukaryotes 
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Joined: Thu Dec 09, 2010 1:10 am
Posts: 4
Post Working with multicellular Eukaryotes
I'm experimenting with mushrooms at the moment. Not necessarily, as everyone assumes, the magic variety.
Psilocybe cubensis is a really interesting organism, and easy enough to grow that it makes a good model, but there are legal issues around its production.
So I've picked Pleurotus ostreatus am experimenting with the best production methods, and getting a microscope after Christmas.

I was wondering whether has succeeded in getting multicellular fungi to express anything interesting?
I enjoyed the thread on mutagenesis, and might try exposing mycelia cultures and spores to UV and then fruit.
I pretty much have my HEPA laminar flow hood functioning, but don't have a fume hood, so I'll avoid the chemical mutagens for the moment.

Does anyone know a mechanism for insertion? The holy grail would be a fluorescent protein, but that's probably really difficult.

Any ideas would be appreciated, and I'll post any interesting results!
C


Fri Dec 24, 2010 1:23 pm
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Joined: Fri Oct 08, 2010 6:08 am
Posts: 58
Post Re: Working with multicellular Eukaryotes
Check out this article:http://findarticles.com/p/articles/mi_m1200/is_n6_v142/ai_12535871/

In terms of insertion, viral vectors seem to be the way to go (although how you will obtain those is another issue). I haven't read very much about fungi, but I think the other methods of transformation (electroporation, chemical transformation, and gold particle bombardment) would work. See http://www.jstor.org/pss/3761368 for more details.


Tue Dec 28, 2010 3:05 pm
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Joined: Thu Aug 19, 2010 3:18 am
Posts: 26
Post Re: Working with multicellular Eukaryotes
UV mutagenesis of spores is very feasible. There should be plenty of papers out there on the subject, particularly if you look up protocols for neurospora crassa, which is a commonly studied fungi.

Genetic Engineering is going to be more challenging, particularly if you are working with a fungi where protocols have not been optimized.

Furthermore, I'm going to tell you what I've told others on this site. Genetic Engineering of any eukaryote typically begins with bacteria. What I mean by that is that the vectors and genes are first grown and manipulated in E. coli before being inserted into the target organism, this is pretty much true regardless of what method you use.

So before you think of Genetically Engineering a fungi, you first need to get a grasp of working with bacteria, transforming them with plasmids, growing them, and isolating the plasmid DNA.

E. coli is simple to work with, but its a necessary first start.


Wed Dec 29, 2010 6:08 pm
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Joined: Thu Dec 09, 2010 1:10 am
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Post Re: Working with multicellular Eukaryotes
Thanks, guys.

chimera's sources give me a lot to go by and chad, I guess you've just told me how I'm probably going to go about it. I don't particularly trust bacteria, and I'm going to have to step up my whole lab, as well and sterile-procedure.

I'm pretty far from having non-trivial results, but I'll be happy to post them when I do.

I googled around with the above search terms, and there are a fair number of papers on inserting genes into filamentous fungi, with electroporation being especially promising.

I guess I have to set up an E. coli lab, now!

I'll be checking in and posting if I have anything to contribute.
Thanks,
C


Thu Dec 30, 2010 1:37 am
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Joined: Fri Oct 08, 2010 6:08 am
Posts: 58
Post Re: Working with multicellular Eukaryotes
About microscopes-Home Science Tools, an educational science store, has some great microscopes for good prices. That's where I got mine from.


Tue Jan 04, 2011 5:58 am
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