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Do you think this is possible ?
    #1605634 -

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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Re: Do you think this is possible ? [Re: Ghengis Khan]
    #1605669 -

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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Re: Do you think this is possible ? [Re: ]
    #1605713 -

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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Re: Do you think this is possible ? [Re: calicyco]
    #1605734 -

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...


--------------------

The blue... the blue!!!

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Re: Do you think this is possible ? [Re: ]
    #1605759 -

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 05:56 PM)

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Re: Do you think this is possible ? [Re: Ghengis Khan]
    #1605863 -

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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Re: Do you think this is possible ? *DELETED* [Re: megosh]
    #1605932 -

Post deleted by poke smot!

Reason for deletion: x


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Re: Do you think this is possible ? [Re: megosh]
    #1605943 -

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


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

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Re: Do you think this is possible ? [Re: micro]
    #1606073 -

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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Re: Do you think this is possible ? [Re: Ghengis Khan]
    #1606139 -

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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Re: Do you think this is possible ? [Re: poke smot!]
    #1606481 -

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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Re: Do you think this is possible ? [Re: Ghengis Khan]
    #1606493 -

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 (06/03/03 10:42 PM)

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Re: Do you think this is possible ? [Re: Ghengis Khan]
    #1607193 -

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


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: Do you think this is possible ? [Re: Joshua]
    #1607364 -

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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Re: Do you think this is possible ? *DELETED* [Re: psilo9com]
    #1607391 -

Post deleted by poke smot!

Reason for deletion: x


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Re: Do you think this is possible ? [Re: poke smot!]
    #1607648 -

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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Post deleted by Papaver [Re: Ghengis Khan]
    #1608575 -


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Re: Do you think this is possible ? [Re: ]
    #1609237 -

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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Post deleted by Papaver [Re: Funger]
    #1609289 -


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Re: Do you think this is possible ? [Re: ]
    #1609809 -

Well said. Agreed.

Interesting conversation though!!!

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Re: Do you think this is possible ? [Re: Funger]
    #1610208 -

What is wrong with liquid mycelium culturing in big aerated tanks?


--------------------
Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Do you think this is possible ? [Re: Anno]
    #1610797 -

I'm not sure if this question was intended to be directed to me, but what the hay...

Doesn't mycelium only produce trace amounts of psilocybin/psilocin? What the author of the thread is trying to achieve is a bacteria that produces the active compounds.


Edited by Funger (06/05/03 07:22 AM)

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Re: Do you think this is possible ? [Re: ]
    #1610800 -

If you want to hammer out what is "right" or "appropriate" about the drug culture, then please do it in a different forum. This forum is for the discussion of advanced cultivation techniques and ideas, and I think that this idea definitely falls under that category.


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Re: Do you think this is possible ? [Re: Funger]
    #1610819 -

>Doesn't mycelium only produce trace amounts of psilocybin/psilocin?

Not that little in my experience.
And you can make up for the minor amounts with bigger amounts of mycelium.


--------------------
Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Do you think this is possible ? [Re: Anno]
    #1612065 -

Just some thoughts:
I forget who said it, but it's not bacteria but yeast that I suggested cloning into. Bacteria's sometimes tricky stuff to play with, gotta be way more sterile. Also, having smelt my fair share of bacterial cultures (cheesy shroom jars/armpits on a hot day, etc...), I"d far rather gulp down some yeast powder/ extract.
Also, like growing cannabis (were it legal) , this would take the power out of the hands of dealers/ cartels/syndicates- who ever heard of a yeast monopoly ???

"What is wrong with liquid mycelium culturing in big aerated tanks?."
Hell, if I could afford some big aerated tanks then I'd already have hired a team of scientists to bioengineer the yeasts. This is a not a shrooms vs. yeast thread I was trying to start- that's rather a lame thing to get all worked up about. Also, it's easier to hide a couple of bottles away in a dark place somewhere, and turn them over every few hours, than concealing a shitload of machinery and gleaming metallic cylinders.... "ummm.. well officer........ those are just errrr... giant fish tanks!"

Some additional side benefits ..... Psychedelic fruit wine!!! Any d.i.y shroom grower
should find this interesting if you don't already know about it:
http://www.wineworldfdw.com/fruit_wine.html
If you can already grow shrooms, this wine- making should be a breeze. However, substitute the word yeast in that article with "Psilo- yeast" (I have to give a name) and it gets a lot more interesting. Imagine sipping a glass of nice apple/ raspberry/peach wine, and tripping........ you'll have to admit it's an appealing idea.





Edited by Ghengis Khan (06/05/03 03:19 PM)

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Post deleted by Papaver [Re: Funger]
    #1612246 -


Edited by paradis (06/05/03 04:25 PM)

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Re: Do you think this is possible ? [Re: ]
    #1612317 -

Relax a little........ my sole purpose in life is not to produce "magic yeast." Other ideas that also occured to me were super enhancing human senses via: increasing the number of scent receptors in the nose, altering the cells at the back of the eyes to percieve infra red and ultra violet, installing a fluid filled cavity in the head to percieve sonar, etc..... inserting THC producing genes into common weeds...... blah blah blah, those are some I can remember. But actually going out doing these things is something different.
In the good old days I used to smoke a lot of weed, grow shrooms, and read a lot of science fiction while studying microbiology- you get some strange ideas with this combo.
But this yeast thing is entirely possible, and WILL be done in the future



Edited by Ghengis Khan (06/05/03 04:48 PM)

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Post deleted by Papaver [Re: Ghengis Khan]
    #1612359 -


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Re: Do you think this is possible ? [Re: ]
    #1612811 -

That's a really good idea :smile:

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Re: Do you think this is possible ? [Re: ]
    #1612813 -

Don't fix it if it ain't broke. What's the difference between 1 gram of shrooms and .03 grams of psilocybin? .97 grams of mushrooms, which are half the fun!

Unless your idea of fun is genetic recombination and hours upon hours of labwork (which I can understand.. :smile: ), then hey - go for it!





--------------------

The blue... the blue!!!

Edited by Rustik (06/05/03 07:41 PM)

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Re: Do you think this is possible ? [Re: Rustik]
    #1614149 -

So that's actually very inefficient Rustik, 97% of the matter is only for 50% of the fun! :wink:

But I agree, just the growing is half the fun, I wouldn't want to miss that.

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Re: Do you think this is possible ? [Re: koraks]
    #1615938 -

Well, if you don't mind losing out on the 'fun' of growing mushrooms, this is actually a rather 'simple' method to make bulk materials -- if you have access to a genetics lab and such, know which genes produce the chemicals, etc. It's used by pharmaceutical companies to produce several differnt types of drugs -- I know that most penicillin is now produced from gengineered E. Coli vats.

If you used a yeast instead of a bacteria, you could have insta-psilobycin anywhere. Packet of yeast, bucket, sugar water and some nutrients, fish tank aerator and a bubbler stone ... 12-24 hours later, you've got ten gallons of psychedelic yeast. It'd be very easy to strain the stuff and dry it. Furthermore, culturing more would be as simple as setting up another bucket and taking a few drops from the first.



--------------------
"Do we want the stars? We can have them. Can we borrow cups of fire from the sun? We can and must and light the world." --"On the Shoulders of Giants", Ray Bradbury

All of my posts are full of fiction and blatant lies.

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Post deleted by Administrator [Re: Ghengis Khan]
    #1619600 -


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Re: Do you think this is possible ? [Re: YellowPurpleHills]
    #1621871 -

I believe it would be possible if proper $ were allocated to research. But it wont be, so it wont. No home lab gonna be able to do DNA splicing, i don't think.


--------------------
Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--

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Re: Do you think this is possible ? [Re: Raadt]
    #1622648 -

Yeah, that money thing, again :wink:

--
Micro


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

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Re: Do you think this is possible ? [Re: micro]
    #1625402 -

Well, yes.  Technically -- for a specialist in recombinant DNA in a fully equipped lab -- it should be pretty easy.  For you or me?  Hell no.  :wink:


--------------------
"Do we want the stars? We can have them. Can we borrow cups of fire from the sun? We can and must and light the world." --"On the Shoulders of Giants", Ray Bradbury

All of my posts are full of fiction and blatant lies.

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Re: Do you think this is possible ? [Re: Sev]
    #1625968 -

Actually, I do work with RNA in a "fully equipped lab."  They pay me, too.  :wink:

--
Micro


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

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Re: Do you think this is possible ? [Re: micro]
    #1626023 -

well then, how about it?

Actually, all this talk, and we've missed the better option. Splice that gene into a Garden Giant! For those unfamiliar, garden giants fruit on a wide range of substrates from wood to straw, the beds fruit for years, and individual mushrooms weigh up to 7 pounds a piece!

So, how about it micro, are we going to see some psychoactive yeasts and garden giants coming out of your lab anytime soon?


--------------------
"From a certain point of view"
-Jedi Master Obi Wan Kenobi

PM me with any cultivation questions.

I just looked at my profile and realized I had a website at one point in time on geocities, it's not there anymore and I have no idea what I had on it. Anybody remember my website from several years aga? PM if so please.

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Re: Do you think this is possible ? [Re: mycofile]
    #1626047 -

Last time I checked the vector kit for s. cervasia (don't know how to spell that) is about $450.  Too much to come out of my pocket :wink:  We don't work with it here, so I'd need a lot of stuff we don't have.  I also don't think they'd appreciate me using their reagents for this.  It's an interesting topic of discussion, though.  Maybe some day when I have my own grant :wink:

As for other types of mushrooms, though, there has actually been some success in splicing hyphal cells' genetics between two types of fungi using polethylene glycols; this would be cheap.  Just don't know the success rate.

BTW -- I WILL do this at some point in my life, just might not be for a few years.  This is simply because it SHOULD be done.  When I do, I'm going to disseminate cultures to random people so that everyone will have them :wink:

On of my life goals.

--
Micro


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

Edited by micro (06/11/03 09:51 AM)

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Re: Do you think this is possible ? [Re: micro]
    #1626133 -

I'M random...  :wink:

a growin original

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Re: Do you think this is possible ? [Re: growin]
    #1626356 -

Quote:
I'M random... 



Apparently so.... :wink:

--
Micro 


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

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Re: Do you think this is possible ? [Re: micro]
    #1637138 -

Quote:
As for other types of mushrooms, though, there has actually been some success in splicing hyphal cells' genetics between two types of fungi using polethylene glycols; this would be cheap. Just don't know the success rate.



Can you elaborate on that a little?


--------------------
"From a certain point of view"
-Jedi Master Obi Wan Kenobi

PM me with any cultivation questions.

I just looked at my profile and realized I had a website at one point in time on geocities, it's not there anymore and I have no idea what I had on it. Anybody remember my website from several years aga? PM if so please.

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Re: Do you think this is possible ? [Re: mycofile]
    #1638283 -

I'm looking at the textbook, I think.... Might take a little bit.

--
Micro


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

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Re: Do you think this is possible ? [Re: micro]
    #1638669 -

Turns out it's actually protoplast fusion, so it isn't as easy as I thought....

I just read the whole damn chapter on genetics, again, and of course I couldn't find it.  Prolly wasn't even the right book :wink:

It made me mad, though, so I did an extensive search, and after a while found this:

http://fungus.org.uk/cv/thesis_fig5.4.htm

and another reference not used in that thesis, if you want (sounds very relevant:)

Ogawa, K. 1993. Interspecific
and intergeneric hybridization of edible mushrooms by protoplast fusion.

While reading the chapter again, however, I came across an interesting piece of info related to mating 2 incompatibale isolates from the same "parental strain."  Take the two, make them each nutritionally deficient for a seperate compound (I guess by breeding or manipulating the genome) and culture on a medium lacking both.  They failed to mention, however, the definion of parental, so I have no idea how much genetic variation is acceptable.

--
Micro


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

Edited by micro (06/16/03 10:41 PM)

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Re: Do you think this is possible ? [Re: micro]
    #1639108 -

Thanks for the research.  I found this
Quote:
Protoplasts isolated from mutant strains were fused using polyethylene glycol


somewhat disheartening.  With no more info other than "mutant strains", who knows what was really going on.

I would love to see the other reference, I guess I'll have to go to the library (30 minutes away :frown: if I want that one...

thanks


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PM me with any cultivation questions.

I just looked at my profile and realized I had a website at one point in time on geocities, it's not there anymore and I have no idea what I had on it. Anybody remember my website from several years aga? PM if so please.

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Re: Do you think this is possible ? [Re: mycofile]
    #1640289 -

"With no more info other than "mutant strains", who knows what was really going on."

Sorry,  I hope this helps:

ABSTRACT

Fundamental genetic analysis of the imperfect fungus Cephalosporium acremonium was undertaken following fusion between protoplasts of nutritionally complementary strains. Protoplasts were obtained in high yields from strains of C. acremonium using the commercial enzymes Cellulase CP or Novozym 234. Other factors affecting protoplast release were investigated to maximise the effectiveness of these enzymes. Optimal conditions for the reversion and fusion of protoplasts were also established.

Haploid recombinants were isolated directly from the fusion plates after plating PEG-treated protoplasts onto a variety of selective media. To minimise any adverse effects caused by the presence of an osmotic stabilizer in the medium a method was developed using cellophane discs to transfer the developing colonies onto normal media as soon as they had become resistant to osmotic shock. Using these techniques mutations at thirteen gene loci were assigned to eight different linkage groups.

The primary colonies obtained on the fusion plates were heterogeneous consisting of haploid segregants and unstable heterozygotes (aneuploids and possible diploids). Overall the results suggest that recombination in C. acremonium occurs by a similar process to that found in other fungi with a parasexual cycle.

This work has demonstrated the value of protoplast fusion methodology in genetic investigations with this fungus. Hybridization between strains of C. acremonium and Cephalosporium chrysogenum was also achieved although these are probably identical species. However, attempts to hybridize the more distantly related species, C. acremonium and Emericellopsis minima, proved unsuccessful.

Note: Cephalosporium acremonium (now known as Acremonium chrysogenum) is used commercially to produce the ?-lactam antibiotic cephalosporin C.

More results:
http://fungus.org.uk/cv/thesis_fig4.1.htm

________

A lot of other stuff can be found on the site that I have yet to read:

http://fungus.org.uk/sitemap.htm

Most of the good stuff is at the bottom :wink:

--
Micro


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