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Offlineleschampignons
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: Alan Rockefeller]
    #20916887 - 12/02/14 12:32 PM (9 years, 3 months ago)

Quote:

Alan Rockefeller said:
Not too hard either, costs about $2000.  The harder part would be to identify which gene codes for the tryptamine pathways.




Is this cost roughly the same for all organisms? Are animals or plants more expensive? Amazing how cheap it is nowadays.


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OfflineAlan RockefellerM
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: filamentous]
    #20917971 - 12/02/14 03:49 PM (9 years, 3 months ago)

Quote:

filamentous said:
Let's get this $2k crowd funded then





I have thought about that, but the major crowdfunding websites have a clause about questionable research, which seems to exclude Psilocybe since the sequencing people won't have a schedule I license.

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InvisibleAmphibolos
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: Alan Rockefeller]
    #20922167 - 12/03/14 11:24 AM (9 years, 3 months ago)

Is the fungal mycelium considered a schedule 1 drug?


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OfflineAlan RockefellerM
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: Amphibolos]
    #20922480 - 12/03/14 12:41 PM (9 years, 3 months ago)

Quote:

Amphibolos said:
Is the fungal mycelium considered a schedule 1 drug?





Yes, if it is dikaryotic.    Monokaryotic is said not to produce psilocybin, so it would not test positive for a controlled substance.

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OfflineMushdiver
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: filamentous]
    #20927394 - 12/04/14 09:54 AM (9 years, 3 months ago)

Quote:

filamentous said:
It won't be profitable?  It's not about profits.  It's about having an easier route to get to psilocin, and getting it faster as to get more people turned on.




If it truly is an easier or faster route to get Psilocybin to people, then it WILL be profitable. This whole argument is moot. Good luck maintaining a stable form of psilocin producing bacteria.

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OfflineKman1898
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: Mushdiver]
    #20927455 - 12/04/14 10:09 AM (9 years, 3 months ago)

Subbed


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Difficulty has more to do with reading abillity and ability to precisely follow directions. You need no knowledge of chemistry whatsoever, you just need to understand some basic principles as simple in concept as: water boils at 100C and freezes at 0C. Otherwise all published syntheses of organic and inorganic compounds can be reproduced successfully by pretty nearly anyone with at least average intelligence. Problems always have to do with availability of materials, not esoteric knowledge.

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Offlinepantsboy
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: Mushdiver]
    #20928378 - 12/04/14 02:00 PM (9 years, 3 months ago)

Quote:

Mushdiver said:
Quote:

filamentous said:
It won't be profitable?  It's not about profits.  It's about having an easier route to get to psilocin, and getting it faster as to get more people turned on.




If it truly is an easier or faster route to get Psilocybin to people, then it WILL be profitable. This whole argument is moot. Good luck maintaining a stable form of psilocin producing bacteria.




you make that sound like it would be a difficult thing.


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InvisibleAmphibolos
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: pantsboy]
    #20928826 - 12/04/14 04:12 PM (9 years, 3 months ago)

You make that sound like it would be an easy thing


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Offlinepantsboy
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: Amphibolos]
    #20928862 - 12/04/14 04:20 PM (9 years, 3 months ago)

if the sequence coding for psilocin were isolated, than yes it would be.
bacterial transfection is cheap and easy. GFP could be used to identify transfected cells and selection markers can be used to positively select for the transfected cells. new colonies would be formed from the transfected cells and over time, you'd end with a culture made entirely of stable, psilocin-producing bacteria. why does that seem hard to do?


--------------------
Acid doesn't hurt when you're on fire. :frown:




"Mushrooms are only similar to penises in their appearance." - LeBron James (2013)

ToiletDuk said:
"Bus squelching is not to be laughed at."

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InvisibleAmphibolos
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: Amphibolos]
    #20928953 - 12/04/14 04:39 PM (9 years, 3 months ago)

Well, first its not "a sequence coding for psilocin", its more like a set of genes (probably arranged in cluster) that code for the enzymes that are used in the pslocin biosynthesis pathway. And no, they are not known yet.

Really simplified method:

So what you'd have to do is  sequence the genome of a psilocybin producing fungi, compile the data, and analyse it to find the wanted sequencces.

Then you use that knowledge to produce the gene sequence individually and introduce them in a plasmid. If they are in a cluster it should be easy, if they are not you have to do a bunch of rounds of PCR.

Also, the sequences will probably contain exons, so you'd have to remove them if you are using a bacteria as host.

Etc, etc.


Its not as easy at it looks, and i probably have forgotten some things. You'd also need alot of founding to do the research, and unless you are backed up by a lab team, doing everything alone might take some time to design. Don't forget that compiling / annoting a genome can be the subject of some Master thesis, so its definitely not an easy task.


However, if we can get a bunch of people to work on the sequence that we got, i'm sure we can work this step out. I'm not that good in bioinformatics, but i'd be interested to learn, so if you set the sequencing of the fungi, i'd be happy to help with this.


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Edited by Amphibolos (12/04/14 04:46 PM)

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Offlinepantsboy
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: Amphibolos]
    #20929155 - 12/04/14 05:22 PM (9 years, 3 months ago)

yeah, well if it wasn't clear i was saying that the transfection and producing a stable germ line would be quite simple in the hypothetical situation that psilocin producing organism had already been sequenced and analyzed.

but on the subject of sequencing and genomic analysis,
a lab right next to me is offering genomic sequencing as low as $150 a day and i understand they sequence around 300 million base pairs per day. it's pretty insane how fast, efficient, and cheap the technology has become. my friend sent a transfected cell line off to them the other day for sequencing.

i think gene prediction is one of the most interesting aspects of genomic annotation. i would love to be able to work with others who are working on genomic analysis of a psychoactive organism.
but i only have a geneticist's understanding of genomic analysis by using knockout genes and other techniques that would never really be applicable to gene finding. i know very little of bioinformatics although i'd love to be trained in the field. my friends who went in to bioinformatics and genomics use a gene prediction software nowadays. the whole process is fast, cheap, and automated. i think the whole gene finding process has been so simplified by modern technology that it is no longer the daunting task that it once was.


--------------------
Acid doesn't hurt when you're on fire. :frown:




"Mushrooms are only similar to penises in their appearance." - LeBron James (2013)

ToiletDuk said:
"Bus squelching is not to be laughed at."

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OfflineVauxhall
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: pantsboy] * 2
    #20964104 - 12/11/14 09:42 PM (9 years, 3 months ago)

Having just finished my Bachelor of Science with a major in Microbiology and Bioinformatics, I can't help but chime in here briefly.

Sequencing tech HAS reduced in cost massively, it is now crazy cheap to get a fragment of DNA sequenced. The problem here wouldn't so much be the sequencing, as creating the genome sequence. These are two very different things. Once we know the genomic sequence, it's easy enough to take a fragment and line it up to that sequence. However, without the genomic sequence we don't know where any given fragment of DNA will line up.

So whoever said that it costs $2000 for a genome sequence, thats for a genome we have already sequenced. For example: We know the Human genome, but if you wanted to know your personal genome then this would be the case.

As it stands currently, it takes whole teams of specialists still to construct genomes from scratch. It involves sequencing the target organisms DNA 30-90 times over and then aligning all the fragments together into one big Genome scaffold.

Gene annotation and identification is then another huge task. Another example, of the 21,000 human genes identified, we know what roughly half of these actually do. The human genome is one of the most heavily studied of all genomes.


If we actually got to the point of having an appropriate genome sequenced, and even identified the genes in question, then it would be a relatively easy task to clone it into E. coli. Given all the data I just suggested and a working lab and reagents I have the skills to do that already, it's not hard work and has been done many times over. The hard part here is the Genome sequence and gene identification and annotation. This is the science of BioInformatics, go look it up if youve any further questions.

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Offlineleschampignons
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: Vauxhall]
    #20964460 - 12/11/14 10:56 PM (9 years, 3 months ago)

Very interesting to read. Thanks for chiming in.


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OfflineKman1898
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Re: Transplanting genes from fungi into bacteria (psilocin related) [Re: leschampignons]
    #20965666 - 12/12/14 08:20 AM (9 years, 3 months ago)

Thanks for the info. Cool to hear you just got your BS too!


--------------------
Difficulty has more to do with reading abillity and ability to precisely follow directions. You need no knowledge of chemistry whatsoever, you just need to understand some basic principles as simple in concept as: water boils at 100C and freezes at 0C. Otherwise all published syntheses of organic and inorganic compounds can be reproduced successfully by pretty nearly anyone with at least average intelligence. Problems always have to do with availability of materials, not esoteric knowledge.

Extras: Filter Print Post Top
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