Author Topic: Not insects! Trees!  (Read 7396 times)

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godshrink

  • Guest
Re: Not insects! Trees!
« Reply #20 on: July 18, 2000, 03:13:00 AM »
John Galt;  glad you have arrived. You and Mr. Smith an d Mr. Green and Rh0dium gonna
bring it on , in my dreams. a new world order.  We know so little. Sure you'll keep that in mind.
Desperate people would trip, if they could afford it. More than 1/2 of the global human producrs
is desperate. Drugs have no chance of changing the world if they aren't free. Swallow this fact,
love bunnies: the average annual ncome. on our lovely planet, is $1000. Much less in places.
Let us not exclude anyone in our dreams of a better world. Sorry about moralizing.
I hate me for this.


john_galt

  • Guest
Re: Not insects! Trees!
« Reply #21 on: July 18, 2000, 08:15:00 PM »
Godshrink, it is time for me to moralize.
Perhaps this should go in a different forum, but you brought it up. World poverty and drug prohibition spring from the same basic moral fallacy, that a person does not have a right to his/her own life. It is precisely that fallacy that must be eradicated from government. Drugs should be free, like free speech, not like free beer, and if trade were free, they would bee. Furthermore, freedom of trade would solve that little poverty problem too.
End Taxes Today: Launder that Drug money in Belize :)
-John Galt


john_galt

  • Guest
Re: Not insects! Trees!
« Reply #22 on: July 18, 2000, 08:37:00 PM »
You have my mind a reelin', Rhodium. Now, do you know the name of the enzyme? I am in a library right now, and am primed for doing some protein searching.

-let there be E on earth, and let it begin with me :)
-John Galt


Beaker

  • Guest
Re: Not insects! Trees!
« Reply #23 on: July 19, 2000, 06:29:00 AM »

Don't they inject yeast with dna all the time in biotech companies?


Yeah, and 99.9999999999999999999999999999% of the time the gene is not expressed properly.


Step 3. This one is hard, you have to get the DNA for the enzyme into the nucleus of the cell. Pay a biochem company to do it for you.


I hope you're rich.

It's a hell of a lot more complicated than just getting the DNA into the nucleous. This technology is about 20 years away from being fully developed. As it now stands, this sort of thing can be done, but is extremely difficult to do in any case, and often can't be made to work in any but the most favorable circumstances. Besides, genetic engineering is only really economical for making complex protein products that are difficult to synthesize in the laboratory, not for making simple, easily synthesizable drugs like MDMA. Super-ergot or Super-opium poppies are more realistic, but you wouldn't need to use genetic engineering techniques, only selective breeding.



godshrink

  • Guest
Re: Not insects! Trees!
« Reply #24 on: July 19, 2000, 03:35:00 PM »
so, i'm gonna keep on analyzing tropical beetles for interesting alkaloids until someone tells me a really good "Eureka" story. For those of you curious about lagrger, cuter creatures, ever wonder how a Koala
can digest eucalyptus leaves and extract everything it needs from them? What on earth is it doing in that stomach? What other beasties live on a diet high in aromatic oils? Best of luck to all of you.


MrGreen

  • Guest
Re: Not insects! Trees!
« Reply #25 on: July 29, 2000, 09:14:00 PM »
Perhaps yeast isn't the right organism. (if it fails 99% of the time as someone said)
Bacteria, however, might be better.  After a weekend of surfing I've found TONS of examples where people put things into bacteria.  (I'm still affectionate towards putting things in the sassy seed due to the fact it contains safrole already.)

Advantages of bacteria:
 Common techniques already in literature (e.g. putting DNA into bacteria, growing conditions, etc)
 Cheap to maintain
 Easy to grow
Disadvantages:
 Would have to extract the product out of bacterial 'soup' (I think these techniques are out there already)
 Would require access to DNA to put into bacteria (e.g. Transaminase enzyme DNA)

Care to add the pro/con list above?

I'd like to know that transaminase enzyme also Rhodium!!

My wish list:
Someone to find enzymes that would help convert dopa to mdma


bizarium

  • Guest
Re: Not insects! Trees!
« Reply #26 on: July 30, 2000, 05:27:00 AM »
Godshrink: I think i can dig where you're comoing from... but to extrapolate on your logic, wouldn't we save time investigating insects that are predatory on the insects that are digesting/concentrating favorite alkaloids? If DDT was the desired substance, we would do well to look far down the line on the food chain
Bugs eat plants, fish eaats bug, bigger fish eats fish, bird eats bigger fish, bear eats bird, and so on? Each organism concentrating chemicals. I've heard that polar bears are so far down the chain Or should i say highup on the chain?) tha eating their livers can bee a toxic experience. Bees! Eat not of polar bear liver!

bizarium

  • Guest
Re: Not insects! Trees!
« Reply #27 on: July 30, 2000, 05:39:00 AM »
one would think, if our human prejudices are correct, that the simpler the organism, the easier we can manipulte it. The aids virus has put some craks in that theory, but still, wouldn't viruses bee the place to manipulate for bio-chemical-engineering? Or, how about prions? If yeasts are gonna bee uppity, why not drop lower?

My personal hunch is that the lowliest creatures are reflecting the loftiest intelligence. Our huuman ego prevents us from seeing the obvious: that our liver flukes are smarter than us. After all is said and done, survival is the only I.Q. test we have. So, how ya doin'?

MrGreen

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Re: Not insects! Trees!
« Reply #28 on: July 30, 2000, 06:41:00 AM »
Regarding prions and viruses... a quick look at any news website tells me prions are just proteins, not organisms, therefore nothing to modify. (they're just a single entity, not a working machine such as yeast is)
As for a virus, I think they're a parasitic species, incabable of 'surviving' without a host.  They don't have enzymes that modify chemicals, i believe.

Don't flame me if i'm wrong, but i think both of these species are out of the question.


MrGreen

  • Guest
Re: Not insects! Trees!
« Reply #29 on: July 30, 2000, 07:08:00 AM »
How 'bout this:
Starting with phenylalanine (an amino acid) and ending in almost-mdma:
C6H5-CH3-CH2-CO2H  -1-> HO-C6H5-CH3-CH2-CO2H -2-> HO-C6H5-CH3-CH2-CO2H -3->
               |                                          |                         |               |
               NH3                                      NH3                    HO              NH3

CH3O-C6H5-CH3-CH2-CO2H -4-> CH3O-C6H5-CH3-CH2-CO2H -5-> CH3O-C6H5-CH3-CH2
          |            |                               |             |                             |             |
        HO          NH3                          HO            NH2CH3                  HO            NH2CH3

Reaction: (enzymes)
1 Phenylalanine hydroxylase
2 Tyrosine hydroxylase
3 Catechol-o-methyltransferase (COMT)
4 Phenethylaminen-n-methyltransferase (PNMT)
5 Aromatic amino acid decarboxylase (AAAD)

Just stick these 5 enzymes into bacteria/sassy tree...

Sooooo close!!
->CH3O- and HO- on ring need joining.
->Maybe that last enzyme (AAAD) is unnecessary.  We really need a -CO2H ---> -CH3 enzyme.


My wish list:
 Someone to find enzymes that would help convert dopa to mdma


bizarium

  • Guest
Re: Not insects! Trees!
« Reply #30 on: July 30, 2000, 05:41:00 PM »
I've heard that many viruses survive without a host; some floating around in the ocean.
Perhaps, some of the compounds we seek could bee the by-products of infection? Do yeasts get viral infections? I wish i knew enough about this to flame. Good luck, Mr. Green.

MrGreen

  • Guest
Re: Not insects! Trees!
« Reply #31 on: July 30, 2000, 06:31:00 PM »
Yes a virus can /survive/ for 1000s of years in the ocean, desert, etc.  But it doesn't /duplicate/ until it gets into a organism.
We need something more complex that will have digestive enzymes, like a transaminase, and big enough to hold maybe 10 new ones.  (see my last post for 5 of them)
From what i see, a virus is just DNA wrapped up in a small package.  The package opens once inside a host cell, and the DNA subverts the host cell machinery (enzymes) into duplicating it. Stupid virus can't even duplicate itself!  Truly remarkable that they've survived/thrived!!  Amazing.

As for a virus infecting yeast, i think it can.  A virus can infect plants and bacteria... i don't see why not yeast. 

Fat_Freddy

  • Guest
Re: Not insects! Trees!
« Reply #32 on: August 07, 2000, 07:37:00 PM »
yes virus' can be used to infect bacteria for our purposes, infact it is one of the main methods used by geneticists and biochemist in genetic engineering.  The basic over view for such a process would be as follows (I think). 

First you would grow the cells of interest (the ones that produce the enzyme of you want, ie rabbit liver enzymes).  Next you would need to create a cDNA library of the cells DNA (a library that only contains the DNA that codes for the expressed enzyme). This is done by liasing (busten them up)your cells  and introducing a pollyT DNA probe (a sequence of DNA that contains only contains thymidine nucleotides)  this will bind to the messenger RNA (RNA that encodes for enzyme expression) from the cells.  Now reverse transcriptase would be added to the mix.  This will bind to the PollyT sequence on the mRNA and use it as a primer and will encode a cDNA copy of the mRNA (have I lost anyone yet).  Ribonuclease H would be added this will degrade the mRNA.  Then Polimerase 1 and ligase will be added wich with out going into it will produce a double stranded DNA copy of all the genes that are expressed in the cell. 

Now we have to introduce this DNA into our plasmid or phage (virus thingies)  this is done by a process known as Terminal transferase I won’t go into it to much as I’m starting to wonder about why I started to write this.  Any way  plasmid vectors are added to the cDNA along with restriction enzymes.   The restrictions enzymes cut up all the DNA (both plasmid and cDNA) in such a way that the cDNA combines with the plamid vector.  This is then added to E. coli or some other bacteria with calcium salts.  The calcium salts make the bacteria permeable to our plasmid vector and vola we have a self replicating library with all the expressed genes from our original cell.

From here you would have to screen the library for the gene of interest using one of many techniques (depending on the gene and what it does you may have to use certain ones)  once you have found the group of bacteria in your library that has the gene you remove it and place it by itself it will the replicate produce a colony that produces shit loads of the enzyme you want.

Of course you could probably just buy these enzymes from companies but when the DEA or what ever restricts their sales those that have their own colonies will have no problems.  

On another note with the above techniques and others it would be quite possible to create bacteria that when placed in safrol could produce MDMA.  Of course the research into creating such bacteria would be expensive (probably) but once a way is found the rest of us could copy the procedure and it would be cheap.  Most of the enzymes, plasmids and probes needed can be bought for not much more than most chemicals.  The best thing would be they would only have to be bought once.  And if the innovation that the hive has applied to them organic chem routes was applied to this we probably wouldn’t have to buy these things as they are already in our blood and lying around in mouldy coffee cups.

Anyway that’s my 2 cents   

Fat_Freddy

  • Guest
Re: Not insects! Trees!
« Reply #33 on: August 07, 2000, 07:49:00 PM »
Something else that’s interesting.  Last week my lecture was talking about green chemistry and how companies were using enzymes to create benzene compounds out of glucose yes GLUCOSE.  If someone created bacteria that turned glucose into MDMA say good bye to the war on drugs.  Talk about turning lead into gold.

placebo

  • Guest
Re: Not insects! Trees!
« Reply #34 on: August 07, 2000, 07:56:00 PM »
SOunds relatively easy actually, surely there is data somewhere on what bacteria/mould/yeast/enzyme/slimey sludge, etc, does what to what molecules!

Then its just a matter of mix and match for our different stages.

RXN 1, bugs eat safrole. blend bugs and extract.
RXN 2, Put yeast into blender.
RXN 3, extract and xtalise.

Simple!


:P <--If I put this here, people think I'm joking instead of a cunt!

Mr_Smith

  • Guest
Re: Not insects! Trees!
« Reply #35 on: August 08, 2000, 03:50:00 AM »
I really think this can be done, but it requires a team effort. You need people with experience in aquiring chemicals, extraction techniques, and sterile technique (someone good at growing 'shrooms). The thing that makes this worth the herculean effort, is that, upon successful completion, MDMA will be about as easy to produce as beer.

An easier "trial run" might be producing GHB/GBL through fermentaion, since there is probably already an organism that can do this on the right substrate.

8)

bizarium

  • Guest
Re: Not insects! Trees!
« Reply #36 on: August 08, 2000, 05:08:00 AM »
Fat Freddy, where have you been? Mr. Smith, can you dig this bent? I'm too stupid to recognize subtle spoof in thes matters. And yet, I can also appreciate Godshrink's angle on this...That with over 1'000'000 species of beetles, there is indeed a good chance that one of them is manufacturing in a way that we like. Hivesters: Any entymologists?

MrGreen

  • Guest
Re: Not insects! Trees!
« Reply #37 on: August 08, 2000, 06:51:00 AM »
Right on Fat_Freddy.  Can you buy the DNA from www.atcc.org  instead of going thru all that work about getting it from cells/transcription/phage blah blah blah.  For example, phenylalanine hydroxylase is at:
   

http://www.atcc.org/phage/cds_p.html


So put this into a bacteria and isolate the colony like you said.  Then repeat with DNA for x,y and z.  Pretty soon you have a bateria that produces e.

Anyone know how you'd extract the 'e' from the bacterial junk ??

foxy2

  • Guest
Re: Not insects! Trees!
« Reply #38 on: August 08, 2000, 10:49:00 AM »
Getting the methyl amine group on there would be the challenge.  I have not heard of one real organism that can add a methyl amine group.  MDA easier because of rabbit livers. Isolating this gene would be a bitch, probably worthy of a several hundred thousand dollar research grant.  Someone call up Paul Allen, he is the only one with the funds and the right politics to make this happen. 


"Vote Libertarian"

MrGreen

  • Guest
Re: Not insects! Trees!
« Reply #39 on: August 08, 2000, 06:55:00 PM »

>Getting the methyl amine group on there would be the challenge.
No challange.  Lots of enzymes do this. (PNMT from my rxn in earlier post.) Most organisms contain *multiple* N-methyl-transferases I've found.

>Isolating this gene would be a bitch, probably worthy of a several hundred thousand dollar.
Isolating has been done already and is comercially available. (PNMT again)

What people don't seem to be catching on to is:
A)biosynthesis costs a moderate amount -once- then virtually nothing ever again.
B)the technology, methods, and skills are available NOW.

If someone new the rabbit transaminase enzyme name (from shulgin/pikhal) we'd be rolling. (i don't have the book and can't find it in his published articles)
Alternatively, if someone new an enzyme that would close the ring structure (my earlier post) we'd also be rolling.  What the hell is this reaction even called?!  (ring closure)

CH3-O-R                         O-\
          |             --->  H2C    R
        HO                         O-/