Re: [DIYbio] Re: Open-Source Enzymes

When I looked up ice nucleation proteins on google patents I found some worrying looking patents. Universities don't usually worry about patents, so I'm not sure how seriously to take the implied suggestion that ice proteins can be "Used by anyone". Still, would value more expert opinion there: are ice proteins unencumbered by patents?

Mac Cowell <mac@diybio.org> wrote:

>Would ice-affinity purification work? You link an anti-freeze protein
>moiety to your protein of interest, express, lyse, and expose the
>lysate to a probe that is slowly and evenly cooled (?) to encourage ice
>nucleation.  At some point afterwards the AFP is cleaved off.
>
>Overview: http://2011.igem.org/Team:Yale/Protein
>
>The take team has a poster with more info.
>
>Mac
>
>231.313.9062 // @100ideas // sent from my rotary phone
>
>On Mar 8, 2012, at 4:45 PM, Darrell Montana <dmontan99669@gmail.com>
>wrote:
>
>> Just a heads up on making restrictions enzymes...   If you are going
>to try this you need to co-express the complementary methylase.  If
>not, the RE will chop up the coli genome and kill the host...
>>
>>
>>
>> On Thu, Mar 8, 2012 at 11:45 AM, Derek <derekja@gmail.com> wrote:
>> We started to put together such a project up here at the University
>of
>> Victoria for an igem team, but ultimately decided to do something
>else
>> because of the difficulty of the purification problem. Taq is pretty
>> easy since it is thermostable, but restriction enzymes need to be
>> purified in some way. Some of the earlier restriction enzymes are off
>> patent now, so depending on what you want that may not be as much of
>> an issue.
>>
>> BTW. not an entirely ethical way of getting your plasmid (and one
>that
>> would likely hit patent restrictions), but one that sometimes works,
>> is to take a commercial enzyme and use it as the source of
>> transformation DNA. Make sure you use high competency cells for this
>> because there's not much DNA there, but as good as the commercial
>> purification strategies are they often leave enough plasmid DNA in
>the
>> mix to transform with. You still need to figure out what antibiotic
>> they used and reverse-engineer the purification strategy.
>>
>> --Derek
>>
>> On Mar 8, 11:11 am, Ethan <argentu...@gmail.com> wrote:
>> > Forgive me if this has already been discussed, but what is the
>prevailing
>> > opinion on open-source enzymes. For doing DIY plasmid engineering,
>the most
>> > costly part is probably the restriction enzymes and ligase. Any
>kind of
>> > genetic manipulation (PCR, etc) gets expensive as a result of
>enzyme
>> > prices. Do you think it would be possible to develop some sort of
>> > open-source production method for useful enzymes (ie. open source
>vectors
>> > for expressing the enzymes and some easy way to purify them)? This
>is all
>> > just speculation, and my research into the subject has been
>limited. My
>> > experience in protein purification is also quite narrow.
>> >
>> > I am aware that Cathal has an open-source plasmid vector in the
>works. If
>> > there is an effort to create open restriction enzymes, it would be
>> > convenient to coordinate them with the restriction sites on the
>plasmid. I
>> > am just looking for input on all aspects of such a project. What
>are the
>> > legal restrictions with gene patents? Do you think that it is
>viable to
>> > have some sort of chromatography set up that is reasonable for a
>large
>> > number of DIYers to have access too for purifying the proteins
>perhaps via
>> > a common tag? What do you think the costs (and benefits) of an
>endeavor
>> > like this might be? Thanks for your thoughts!
>> >
>> > -Ethan
>>
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