So, it can be combined with LacY from IGEM in a plasmid and transduced in Salmonella. But how to make the plasmid more stable with this huge insert? I know, what exist some stable big plasmids, what encode pathogenity factors or pathogenity "enhancers" at some bacteria (pPCP1, pMT1 in Y. pestis, pXO1, pXO2 in B. anthracis), but what makes them stable?
Am Dienstag, 26. Mai 2015 23:41:37 UTC+3 schrieb Nathan McCorkle:
-- Am Dienstag, 26. Mai 2015 23:41:37 UTC+3 schrieb Nathan McCorkle:
Looks complete from a length perspective (gene is 1024 AAs, 3 nucleotides per residue, 3607 - 539==1022.66 which is close enough to 1024... Assuming 1024 includes both regions).
What about this plasmid: https://www.addgene.org/13422/ ? It seems, what it has complete LacZ, doesn't it?--
Am Dienstag, 26. Mai 2015 22:02:48 UTC+3 schrieb Nathan McCorkle:On Tue, May 26, 2015 at 11:09 AM, Dorif <dor...@gmail.com> wrote:
> Well, virtually I don't need entire operon for this. It is enought to insert
> LacZ and LacY. If LacZ is functional with deletion in it - it is reaaly good
> idea to use this "cutted down" LacZ to minimize the insert size.
I think Matt's point was that in common plasmids, the lacZ is not
complete... this is for alpha-complementation, since the lacZ gene
product (the protein) is a tetramer composed of two dimers, one of
which is alpha and the other omega... so these common plasmids usually
lack one half of the lacZ gene (which I'm guessing is actually
polycistronic, e.g. two genes in one).
See here for a nice paragraph and graphic of the protein subunits:
http://www.ncbi.nlm.nih.gov/pubmed/23011886
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