Re: [DIYbio] Re: Anybody has E.coli lactose operon in a plasmid?

Two issues.  #1 large inserts in plasmids are typically genetically unstable (due to copy number / potential for rearrangement or deletion). Coli and salmonella genomes will not recombine unless you use special "mutator" salmonella strain.   In any case you are quickly stepping into bacterial genetics PhD program level manipulations. (My background). Chances for success are low, but not impossible. Risk of much time wasted though is very high.  >matt    Dorif <dor...@gmail.com> wrote:    

I'll place it in a plasmid and transform Salmonella with it. If plasmid will be too large - I'll use electroporation. And, as I mentioned before - it is not pathogenic to humans.

Am Dienstag, 26. Mai 2015 19:13:40 UTC+3 schrieb Nathan McCorkle:

On Mon, May 25, 2015 at 10:56 PM, Dorif <dor...@gmail.com> wrote:
> In my country it is not the easiest way, because at us PCR is not well
> developed.(

In that case, what will you do with the operon once you get it? Try to
induce horizontal gene transfer? I wouldn't try that with potentially
pathogenic samples... get some kind of stripped-down plasmid with
otherwise non-scary genes on it... you don't want dead/half-dead rats
to become a carrier and spreading genes that aren't helpful for
humans.

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