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OfflinePTreeDish
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Registered: 04/22/18
Posts: 353
Last seen: 3 months, 14 days
First Genetic Transformation
    #27226598 - 02/25/21 08:49 PM (2 years, 10 months ago)

I'm learning how to genetically engineer filamentous fungi. A key step in the process is transforming E. coli with a DNA plasmid (a circular piece of DNA with your custom genes). If you grow a bunch of E. coli with your genes, well, it will clone your DNA for you that you can then filter out and use to transform other things, like fungi.

Here's some E. coli I recently transformed with the pGREEN plasmid containing an antibiotic-resistance gene and a modified GFP (green fluorescent protein).



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OfflineIce9
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Registered: 03/20/14
Posts: 11,225
Loc: daterapeville,USA
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Re: First Genetic Transformation [Re: PTreeDish]
    #27229516 - 02/27/21 05:09 PM (2 years, 10 months ago)

Eukaryotes do not uptake plasmids.  Works great on prokaryotes, whole nother ball game with Eukaryotes.

Apparently I'm wrong.  Here is a link to an older article discussing the difficulties of getting filamentous Fungi to uptake foreign DNA and express it.


Edited by Ice9 (02/27/21 05:13 PM)


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OfflinePTreeDish
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Registered: 04/22/18
Posts: 353
Last seen: 3 months, 14 days
Re: First Genetic Transformation [Re: Ice9]
    #27229651 - 02/27/21 06:45 PM (2 years, 10 months ago)

Quote:

Ice9 said:
Eukaryotes do not uptake plasmids.  Works great on prokaryotes, whole nother ball game with Eukaryotes.

Apparently I'm wrong.  Here is a link to an older article discussing the difficulties of getting filamentous Fungi to uptake foreign DNA and express it.



Transformation isn't as easy but it's been done many times over the last 30 years. This year alone there have been about half dozen or more papers on CRISPR in filamentous fungi. Here lately, there's been a rise in interest and research into transforming fruit-bearing Basidiomycota.

I'll personally be using electroporation and agrobacterium-based transformation methods, both which have been used successfully on species like Agaricus bisporus. To my knowledge, there are no documented attempts at transforming psychedelics in species like Psilocybe.


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