Re: [DIYbio] How is Agrobacterium Tumefaciens grown? Is it possible to find in soil?

[Cory Tobin]

> This wonderful 'stuff' seems to be the 'gold dust' for doing genetics with:)
> Does anybody know anything about it? Like where to find it, what conditions
> and so on it likes.....How to encourage it?

You can actually grow it on LB just like E. coli, although if you put
it in a 37C incubator it will die. It grows best on LB at 27C.


> Would love to know, even have somebody mail me a sample...maybe:)

If you're in California then you're in luck. I can send you a sample
no problem. If you're in the US but outside California, get a permit
for interstate transportation of plant pathogens here:
http://www.aphis.usda.gov/plant_health/permits/organism/plant_pathogens.shtml
and I'll send a sample. If you're outside the US, check the
regulations in your country and get the proper permits.

Summary: transporting plant pathogens across political boundaries is
complicated, even if it's a "disarmed" strain like GV3101. Your best
bet is to find a plant genetics lab at a university near you and ask
the professor for a sample.


-cory

[Matt DiLeo]

It's a naturally occurring pathogen, causing crown gall among other diseases, and there's plenty of information out in the plant pathology literature on where to find, identify and cultivate it - but you'll want a disarmed lab strain that doesn't causes disease.

[Cathal Garvey]

It's not only the fact that lab strains don't cause disease; it's ease
of DNA manipulation.

Wild Agrobacterium plasmid is a complex mess of regulatory and transfer
sequences, and disease-causing DNA. The lab strains don't just have less
disease DNA, they also separate the transferring DNA from the "payload"
DNA, so there are actually two plasmids in the cell; one for the desired
plant DNA, another for sending the first via an infective pilus.

Definitely don't waste time trying to culture and re-hack a wild crown
gall bug; get your hands on disarmed and re-engineered lab strains and
you'll save a lot of time/money.

--
www.indiebiotech.com
twitter.com/onetruecathal
joindiaspora.com/u/cathalgarvey
PGP Public Key: http://bit.ly/CathalGKey

[Giovanni Lostumbo]

Agrobacterium isn't the only plasmid-transferring microbe, there are ones that aren't gall forming, such as  Sinorhizobium meliloti (http://www.patentlens.net/daisy/bioforge_transbacter/3790/version/default/part/AttachmentData/data/Gene%20Transfer%20to%20Plants.taf.pdf) and Mesorhizobium loti:  http://www.patentlens.net/daisy/bioforge_transbacter/3586.html

[Giovanni Lostumbo]

i also wanted to mention that the alternative transformation techniques use other patents outside of the Agrobacterium patent tree, though that's another long discussion.

[Cathal Garvey]

I hope you're not suggesting importing a _wild_ strain of agrobacterium into the US? That'd be morally and legally a 'bad thing'.

Barra Darcy <bara...@gmail.com> wrote:

>hmmm...
>I live in Ireland but would be going over to Indiana in August, I don't
>
>know if that's any help...it'd leave all the importation ('smuggling')
>up
>to me. I could send you the address if you have a e-mail?
>Is it only found over there or is this world-wide anyway?

>--
>You received this message because you are subscribed to the Google
>Groups "DIYbio" group.
>To view this discussion on the web visit
>https://groups.google.com/d/msg/diybio/-/-0dT21pw_-UJ.
>To post to this group, send email to diy...@googlegroups.com.
>To unsubscribe from this group, send email to
>diybio+un...@googlegroups.com.
>For more options, visit this group at
>http://groups.google.com/group/diybio?hl=en.

--
Sent from K-9 Mail on Android

[Dan Wright]

I see a body cavity search in his future. I think he is trying to get it in the us and bring it back to Ireland. Now you know who to blame for the next potato famine.

[Georgi Dimitrov]

I also wont some, and I can pay .

please let me know if someone haves.

I work on project for life  extension  .

regards 

Georgi

On Sunday, June 17, 2012 5:28:56 PM UTC+2, Barra Darcy wrote:

This wonderful 'stuff' seems to be the 'gold dust' for doing genetics with:) Does anybody know anything about it? Like where to find it, what conditions and so on it likes.....How to encourage it?

[Sebastian S. Cocioba]

If you go outside and see a growth on a branch if a tree, thats mist likely agrobacterium tum. You can do a potato assay and rub inner sections of sliced up tumor growth onto a sterilized potato slice (red russet works best) in a week or two small tumors will form. From there you can rub the potato tumor inner sections on an agar plate with YEP medium. If all works out, that is a wild type strain of agro. From there you can proceed to disarm the Ti plasmid (tumor inducing plasmid) using restriction endos and phage ligase. Then the rest is up to you and your resources. To make agro virulent so it can transfer dna to dicot host, you need 100uM acetosyringone for a certain time period. For plasmid uptake, heat shock works very well. Good luck and please be safe! Happy hacking!

Sebastian S Cocioba

CEO & Founder

New York Botanics, LLC

Sent via Mobile E-Mail 

--

You received this message because you are subscribed to the Google Groups "DIYbio" group.
To post to this group, send email to diy...@googlegroups.com.
To unsubscribe from this group, send email to diybio+un...@googlegroups.com.

To view this discussion on the web visit https://groups.google.com/d/msg/diybio/-/HlwOz0QogR8J.
For more options, visit https://groups.google.com/groups/opt_out.
 
 

[Cathal Garvey]

I'm curious, how does A.tumefasciens relate to life extension?
In theory at least, it can only transform plant tissues?

Regardless, I think it can be cultured on Lysogeny Broth agar (LB) like
E.coli.

On 22/08/12 23:07, Georgi Dimitrov wrote:
> I also wont some, and I can pay .
> please let me know if someone haves.
> I work on project for life extension .
> regards
> Georgi
>
> On Sunday, June 17, 2012 5:28:56 PM UTC+2, Barra Darcy wrote:
>>

>> This wonderful 'stuff'<http://en.wikipedia.org/wiki/Agrobacterium_tumefaciens>seems to be the 'gold dust' for doing genetics with:) Does anybody know

>> anything about it? Like where to find it, what conditions and so on it
>> likes.....How to encourage it?
>>

>> Would love to know, even have somebody mail me a sample...maybe<javascript:>
>> :)

[Mega]

No, it can also transform animal cells and yeast. I googled it up! (But maybe just under specific lab conditions and chemicals??)


You could isolate it from a tree very difficultly, or you simply get one from here. http://www.pgreen.ac.uk/
They haven't  answered my request since one week though. Maybe they need some time.

[Georgi Dimitrov]

thanks for the info , safety first :)

If I have actual progress I will be happy to discuss it here guys 

thanks again and may the force be with you ;)

--
You received this message because you are subscribed to the Google Groups "DIYbio" group.

To post to this group, send email to diy...@googlegroups.com.
To unsubscribe from this group, send email to diybio+un...@googlegroups.com.

To view this discussion on the web visit https://groups.google.com/d/msg/diybio/-/9woJBrff62kJ.

[Sebastian S. Cocioba]

Its kinda interesting that some of the cytokinins used in plants to stimulate cell division and eventually tumor growth has been found in human dermal growths and cancers. Another reason why agro is such a valuable tool in cancer research. :)

Sebastian S Cocioba

CEO & Founder

New York Botanics, LLC

Sent via Mobile E-Mail