The first thing you need to know that it has been done, at least in
tobbaco.
See here: http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0015461
(open access)
The basic idea was somewhat as outlined here - get the lux operon, add
flanking regions of chloroplast DNA, shoot the DNA in, and select for
positives. While one can think of different ways to do it, this is
certainly the simplest. It does require a) a gun, b) the sequence of
the plant chloroplast DNA.
Given this, I don't see a reason why this shouldn't work for any other
plant. As you may have already understood, transforming plants is
somewhat of an art, and if you want to venture away from the commonly
used ones, there is going to be a lot of trial and error. The
particular hormones and conditions of regeneration can vary to a large
extent, and some just won't regenerate (God knows I tried).
Lichens is an interesting but not so well investigated system.
Assuming it is a cyano-fungi (and not algea-fungi) lichen, it might
work. You will probably need a plasmid which is compatible with cyano,
as the e.coli ones are unlikely to work. There are plasmids out there,
but this means sub-cloning. Selection is bound be pretty hard on such
a slow growing organism, though.
How about green algea? There is bound to be some chlamydomonas
swimming happily in a pond in a park near you. They should be PEG
transformable. My guess is that the e.coli plasmid by itself wouldn't
replicate well in the chlamy chloroplast, so your best bet is to aim
for chromosome integration as in the plos one paper.
Idan
On Nov 7, 4:11 am, Mega <masterstorm...@gmail.com> wrote:
> Ok, what I learned so far...
>
> Firstl, it's no big deal for a pro to insert a lux operon containing
> plasmide in an e.coli cell to make it glow. the antibioticum
> ressistance and lacZ' will make it selectable.
> Usuall, there's no more quorum sensing because the have cut it out.
>
> - ''Just'' insert the plasmide into a bacterium and it glows.
>
> To engineer a plant on the other hand is a much more difficult issue.
>
> But what about a path in the middle (don't know if this term is used
> in english ;) )?
>
> Lichens consist of a fungus and canobacterium.
>
> So, i take a plasmide that contains lux and ampecillin resistance.
>
> Now i transform very small pieces of the lichen. Some of the
> Cyanobacteria will be transformed and ressistant to ampecilline.
>
> The fungus should either be ressistant to ampecilline on its own
> (true??) or the cyano spread the ampecilline-degenerating substance
> within the whole lichen.
>
> Polyethleneglycol will help the endoctose to take place...
>
> Then I'll grow them on a Lamp plate to select the lichens that include
> ressistance. Of the bacteria inside the lichen, the ones containing
> the most ampecilline-degenerating substances, these will grow fastest
> and so they will mitose much more often.
>
> I think that could be a way to go?
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