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OfflineGhengis Khan
Kali Baba

Registered: 01/02/03
Posts: 17
Loc: taiwan
Last seen: 13 years, 23 days
Do you think this is possible ?
    #1605634 - 06/03/03 07:19 PM (13 years, 6 months ago)

OK... this one's for the more scientifically minded out there who know a little genetics/ biotechnology.... it's not exactly a growth question but I don't know where else to post

In one of our biochemistry textbooks at university, we had this one example where tryptophan is converted to psilocybin via 2 or 3 enzyme mediated steps (this was 2 or 3 years ago, so the details are a bit vague)- overall, one of the simpler biochemical reactions. Wouldn't it be easy (relatively speaking) to isolate these proteins, sequence them, convert the amino acid code to dna, and use the various methods available to isolate the genes responsible for psilocybin production ?

These could then be easily cloned into, for example, yeast- which is a hell of a lot easier to cultivate than shrooms. Seems to me that if someone actually went to the trouble of actually doing this then psilocybin production could become as easy as making moonshine and, well, the world would be a better place

:grin:

or maybe I just smoked too much before class....

Any thoughts from someone with more experience in this field than me ? genetics was never my strong point. 


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Anonymous

Re: Do you think this is possible ? [Re: Ghengis Khan]
    #1605669 - 06/03/03 07:26 PM (13 years, 6 months ago)

Kind of like drawing a womans pussy onto a watermelon and going at it. WHY would you want to. Growing the mushrooms is most of the fun. Psilocybin and PSilocin can already be produced in a lab, but it is ILLEGAL. Hence the reason this stuff hasn't been done already.


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Invisiblecalicyco
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Registered: 05/03/03
Posts: 355
Re: Do you think this is possible ? [Re: ]
    #1605713 - 06/03/03 07:37 PM (13 years, 6 months ago)

This could totally be done. Questions is.... do you have the funding of a major pharmaceutical lab to genetically engineer bacteria or yeast to produce these compounds?

Its not cheap! An endeavor like this would only make sense if the protein in question were extremely commercially viable. This would take many millions of dollars and several years to get working properly, but once accomplished would make the production of large quantities of psilocybin real easy for a pharmaceutical company.


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OfflineRustik
Where am I?

Registered: 04/18/03
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Re: Do you think this is possible ? [Re: calicyco]
    #1605734 - 06/03/03 07:43 PM (13 years, 6 months ago)

Quote:

Wouldn't it be easy (relatively speaking) to isolate these proteins, sequence them, convert the amino acid code to dna, and use the various methods available to isolate the genes responsible for psilocybin production ?




Well, isolating the protiens would be easy. Everything after that in that sentence would be extremely expensive, time consuming, and would require a shitload of lab/scientific expertise, not to mention equipment.

It doesn't sound "relatively easy," imo...


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OfflineGhengis Khan
Kali Baba

Registered: 01/02/03
Posts: 17
Loc: taiwan
Last seen: 13 years, 23 days
Re: Do you think this is possible ? [Re: ]
    #1605759 - 06/03/03 07:50 PM (13 years, 6 months ago)

mmmmmmmm...... red juicy watermelon.... so soft and warm...

but seriously, I've grown more than my fair share of shrooms, and I loved every minute of it- I know psilocybin can be synthesized in a lab (duh!), but what I'm trying to say is that instead of going through some shitty hours long error- prone chemical synthesis, it would be much easier to get some sterile sugar solution, inoculate it with "special" yeast, leave the solution overnight in a warm dark place, and trip the next day... much cheaper, and much quicker

Also- I know it's expensive, time consuming, etc.. but I've got friends in postgraduate studies already sequencing proteins, and all the rest. Even when I was a student we inserted some simple prefabricated genes into bacteria and yeast..... if you're a doctoral microbiology/ genetics student with access to a well equipped lab, some free time, and some shrooms, you could probably pull this off


Edited by Ghengis Khan (06/03/03 07:56 PM)


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Offlinemegosh
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Registered: 05/03/03
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Re: Do you think this is possible ? [Re: Ghengis Khan]
    #1605863 - 06/03/03 08:19 PM (13 years, 6 months ago)

i dont have much to say here except that your idea is brilliant. so go ahead go back to school get a lab and do it already!


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Invisiblepoke smot!
floccinocci floofinator
Male

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Re: Do you think this is possible ? *DELETED* [Re: megosh]
    #1605932 - 06/03/03 08:36 PM (13 years, 6 months ago)

Post deleted by poke smot!

Reason for deletion: x



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Invisiblemicro
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Re: Do you think this is possible ? [Re: megosh]
    #1605943 - 06/03/03 08:38 PM (13 years, 6 months ago)

Anything is possible! I would think it would be extremely difficult to vector it into something like e. coli, but you prolly could do it relatively easily (notice I said "relatively" and I never said "cheap") into something else that produces aromatic compounds via the shikimic (sic) acid pathway (what you were talking about;) it converts chromisate (eventually) to tryptophan, and then tryptophan -> tryptamine via decarboxylation and then takes off from there, producing whatever is required. I think only plants and fungi fall into this category, however. It's interesting information, but I don't know if anyone has actually ever done this work for the psilocybes (I haven't found any info) and to do everything from scratch would cost a lot of money (primers, vectors, assays, blots, etc, etc.)

Most certainly a good idea to toy with, however.

--
Micro


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(Avatar is Maxxy, a character by Mizzyam, RIP)


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OfflineGhengis Khan
Kali Baba

Registered: 01/02/03
Posts: 17
Loc: taiwan
Last seen: 13 years, 23 days
Re: Do you think this is possible ? [Re: micro]
    #1606073 - 06/03/03 09:34 PM (13 years, 6 months ago)

Thanx micro, it's been a long time since I heard the word "shikimic"- damnit, but I hated biochemistry

I did a google search, and found out that this has already been thought of:
http://www.mescaline.com/misc/

some noteworthy quotes:

"With such a recombinant yeast (and I chose yeast over e. coli due the simplicity of yeast culture, i.e., bread and brewing techniques considered-- and also due to the total lack of pathogenicity of cervesia yeasts in general and the fact that yeasts are eukaryotic"

"all an untrained layman would need to produce these compounds in a pure yield would be: one transformed yeast cell, a bucket, warm water, sugar, amino acids and 12 hours-- and simple acid base extraction techniques to separate the pure psychedelic compounds from the waste materials"

All in all, a very interesting article- I recommend it to anyone who finds the idea appealing, and who understands the science

here's the process more clearly summarized:

1. extraction of active mrna from fungus or cactus

2. cloning of mrna into cdna clones or possible use of mrna/cdna hybrid strands

3. attachment of synthetic linkers to cdna fragments

4. methylation ofcdna fragments

5. restriction of plasmid using sites corresponding to synthetic linkers

6. ligation of plasmid and vector dna using T4 ligase

7. transformation of e. coli pools with ligation plasmid and identification of marker genes on plasmid (typically antibiotic resistance)

8. subdividision of e. coli pools until single unknown genes are characterized by expression products

9. lysis of expanded pools

10. extraction and restriction of plasmids and agarose gel separation of fragments

11. ligation of characterized fragments to yeast plasmid

12. sequential transformation of yeast cells (see references)

13. selection of transformed yeast clones

All quite routine if you have the means and the knowhow. Anyway , it's the idea I find interesting...... you never know, maybe in 10 years time we'll all be logging on to the "Yeastery.org"






















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OfflineJohnnyRespect
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Registered: 04/17/03
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Re: Do you think this is possible ? [Re: Ghengis Khan]
    #1606139 - 06/03/03 10:08 PM (13 years, 6 months ago)

I can see numerous advantages to making psilocybian in the lab. First, no more "guessing" as to potency. second, instead of having dried, bluish mushrooms with you when you went (airplane, concert, in your car, etc), you'd have pills that you could press to look like whatever you wanted (or gelcaps filled with psilocybin). Also, instead of growing mushrooms that psilocybian constitutes only a small aprt of, you could manufacture a few pounds of psilocybn at oen time, and have it forever.

Jr


--------------------
As I felt the soft cool mud squish between my toes, I thought, Man, these are not very good shoes!


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Offlinepsilo9com
Koh Koh Gadget

Registered: 05/20/03
Posts: 267
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Re: Do you think this is possible ? [Re: poke smot!]
    #1606481 - 06/04/03 12:26 AM (13 years, 6 months ago)

Quote:

Kinda like the one guy who made THC oranges. Florida was all @#%!$%#$^!#$^!#$^@#$^ because if they spread, one would not be able to differentiate between regular and THC oranges.... OHHHH damn THC oranges would be the SHIET! I feel like indulging in an orange right now! 




never heard of that but found http://cannabisodling.1av10.nu/orange_high.html
after a google.

Awesome stuff, wish I had a few :smile: 


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Invisiblemicro
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Registered: 05/09/03
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Re: Do you think this is possible ? [Re: Ghengis Khan]
    #1606493 - 06/04/03 12:31 AM (13 years, 6 months ago)

Lol!  Yeastery?  People might get this confuzed with the  Brewery ....

Interesting article!  Just one thing:

Quote:

Also enzymes almost always produce a single enantiomeric species, though that is not a consideration here. 




I don't understand; I guess he did consider that because he talks about it later in the paper (about mescaline.)  Yes, if enzymes are needed which are not present they will have to be inserted, too, but I wouldn't think this would be too hard to overcome; why not just fuck up the other products of the patway that are naturally produced by the host; 99% of the time these secondary metabolites aren't even neccesary.  You would, however, still need to buy vectors, primers, enzymes (restriction, polymerase, etc,) DNA ladder (sounds stupid, but it costs $$$,) sodium azide (you won't find this in your local drugstore,) assaying equipment (for the missing enzymes,) and you'll prolly need to run some blots, northern or southern; whichever way you want to swing it, which means you'll want acrylamide in addition to agarose, temed, 2ME, etc, oh -- a DNA binding matrix (~ $100) so you can extract it out of the cells, CTAB wouldn't hurt, either, unless you want to buy one of those waste-of-money $200 DNA extraction kits,  etc, etc, etc....

Sorry for the rant -- too much speed.

I've noticed there is no such thing as a cheap lab procedure.  If I made as much money as I spend on reagents at work I would never have to work again :wink:

It's a good idea, though, and would work; no reason it wouldn't....  just first the full metabolic pathway must be determined in each organism and linked to the DNA (by cDNA or mRNA or whichever way you want to look at it.)  Wouldn't it be nice if someone had already done this?  If that were the case you could do it pretty cheap.  Also -- S. Cervasia (sic -- I never get that one right) was a good idea; it has already been mapped out; the whole genome, so if you were to use this you would be chopping off half of that work, and could prolly get some shortcuts if you were very, very careful and used small primers.

I have a good breakdown of the Shikimic pathway relative up to Tryptamine if you want it.  I think I got it from Erowid, but I don't remember....

Only one more thing, sorry but I have to:

Quote:

"all an untrained layman would need to produce these compounds in a pure yield would be: one transformed yeast cell, a bucket, warm water, sugar, amino acids and 12 hours-- and simple acid base extraction techniques to separate the pure psychedelic compounds from the waste materials"




??? I take it he's being facetious  :grin: 

I just thought the one cell and bucket things were funny.  I see his point, though.

Just my opinion...

--
Micro

Oh, P.S.

Quote:

or maybe I just smoked too much before class....




So?

hehehehe

P.P.S.

(the speed again -- I'll never go to sleep)  With regard to the THC oranges, my friend made a good point: it would prolly be better using peanuts or avocadoes since they have fat in which the TCH could readily dissolve at higher concentrations without causing a problem.  Makes sense, I guess....

Oh, well.


--------------------
Any research paper or book for free
(Avatar is Maxxy, a character by Mizzyam, RIP)


Edited by micro (06/04/03 12:42 AM)


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InvisibleJoshua
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Registered: 10/27/98
Posts: 5,389
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Re: Do you think this is possible ? [Re: Ghengis Khan]
    #1607193 - 06/04/03 06:53 AM (13 years, 6 months ago)

You guys sound like you know what you are talking about.

I'd like you to consider the biomass conversion of nutrients/carbon between P. cubensis and S. cerevisiae.

I'd also suggest a preliminary test comparing the growth of such a yeast or bacteria in the presence of psilocybin/psilocin.

I think what you talk about is possible. I have already done it in my minds eye.

Call me crazy...but I prefer lazy.

Joshua


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OfflineGhengis Khan
Kali Baba

Registered: 01/02/03
Posts: 17
Loc: taiwan
Last seen: 13 years, 23 days
Re: Do you think this is possible ? [Re: Joshua]
    #1607364 - 06/04/03 10:18 AM (13 years, 6 months ago)

Sigh.......

I've been stuck in the far east for the past 2 1/2 years, thus the memories of my studies have faded quite considerably, and back in the old homeland (south africa), job prospects in scientific fields are bad at best.... my point is that I'm not in any position to do much about this idea by myself- I just had to throw the idea out there b4 my brain atrophies totally- hopefully someone who reads this can bury this deep inside till they have the means to carry it out.

however, it's still a cool idea- thanx to all those who spared some time to give it a thought, and IT CAN BE DONE !!!
Anyway, enough whining:

Joshua- You're not crazy..... you're efficient !! As for your questions, the phrasing makes them unclear-
" consider the biomass conversion of nutrients/carbon between P. cubensis and S. cerevisiae." this can either mean:
1. compare the biosynthetic yields of metabolic products between the two under identical conditions or....
2. Chop up shrooms, feed them to yeast, and see if the yeast produces any trippy chemicals (has anyone tried **shroom beer** ????)

your 2nd question sounds like nr.2 above- I reckon the yeast will just treat the psilocybin like anything else and metabolize it, at the most- doubt if the poor little buggers will trip/ do the tango/grow as big as golfballs, etc... though it would be interesting to see if psilocybin has antimicrobial properties (prob. been done)

Micro: Damm! If anyone can do this it's you!! Seems like all this stuff is still fresh in your brain..... I assume you're studying this shit like I was (microbiology degree), so good luck, and don't waste your degree like I have

THE SPIRIT OF THE YEAST WILL NEVER DIE !!!!!!...... at least not in the beer I'm drinking now

  :blush: 


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Invisiblepoke smot!
floccinocci floofinator
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Re: Do you think this is possible ? *DELETED* [Re: psilo9com]
    #1607391 - 06/04/03 10:43 AM (13 years, 6 months ago)

Post deleted by poke smot!

Reason for deletion: x



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Invisiblekoraks
Registered: 06/02/03
Posts: 25,082
Re: Do you think this is possible ? [Re: poke smot!]
    #1607648 - 06/04/03 01:03 PM (13 years, 6 months ago)

Ok, so technically, it can be done. Now for the more practical aspect of things.
I see two possible ways to get this to work and to get this technique accessible to, say, everyone on this forum.

1. Get your university friends to start the project (some master thesis that would be, eh?), and get them to take a sample of the modified yeast/bacteria home where it can be cultivated and multiplied.

2. Same method, but it requires friends working for a pharmaceutic company. Get them to start the project, and again, let them smuggle some of the modified lifeform into the real world.

One a successfully modified yeast is in your hands, the problem is solved, it seems to me. The difficulty lies in finding people who are willing to start such a project, and consecutively smuggle some of the product outside their institute. Which will of course react in a mercyless manner to the smuggling researches if they ever find out what happened (and that's very, very likely!).

So, however technically it can probably be done, in practice it won;t be easy to find the right people willing to get involved in this process. You're best bet seems to me the university clan, as procedures are considerably looser than in pharmaceutical companies.

If you ever succeed, let me know! I'd love to get my hands on some psilo generating yeast :smile:


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Anonymous

Post deleted by Papaver [Re: Ghengis Khan]
    #1608575 - 06/04/03 06:19 PM (13 years, 6 months ago)



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OfflineFunger
MutantApendanges

Registered: 08/20/02
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Re: Do you think this is possible ? [Re: ]
    #1609237 - 06/04/03 10:20 PM (13 years, 6 months ago)

The point, is that once the initial work is done, mass production would be a breeze.
Negative input when you have no idea of what you are talking about is just a waste of everyone's time.


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Anonymous

Post deleted by Papaver [Re: Funger]
    #1609289 - 06/04/03 10:37 PM (13 years, 6 months ago)



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Anonymous

Re: Do you think this is possible ? [Re: ]
    #1609809 - 06/05/03 01:34 AM (13 years, 6 months ago)

Well said. Agreed.

Interesting conversation though!!!


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