Re: [DIYbio] Craig Venter synthesis, possible with pcr?

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To answer that, you'd have to dig into the origins of the most
commonly used Ampicillin cassette. I believe that's "bla", for "Beta
lactamase", from a Shewanella species, although in the literature it
might have been called "Escherichia" at the time as I believe
Shewanella is a fairly recent reclassification.

One easy route is to take an "old" version of Amp from an early
cloning plasmid and run it through BLAST, selecting bacterial genomes
to filter out cloning plasmids and constructs. First few natural hits
should contain wild species with homologous genes, and you can analyse
their promoters to see if they are dumb/constitutive or if they are
induced by beta-lactam antibiotics like Amp.

Even if they are though, it might be (probably is) an indirect
induction mechanism, something like:
Amp -> Reduced Indicator of Health -> Stress Response Genes -> bla

..so it mightn't be particularly useful, given how many ways there are
to accidentally activate stress response genes during normal culture
conditions.

I'd say that's a shortcut that would require a lot of work to attempt
(characterising Amp-inducible promoters), and may not be worth it,
considering how many well-characterised and effective promoters there
are available in the literature. Use something lazy like light
induction, chemical induction, etc.

On 04/09/2013 02:18 PM, Koeng wrote:


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