Endy:Screening plasmid/Inverter characterization: Difference between revisions
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#Stormo pmid=8445649 | #Stormo pmid=8445649 | ||
#Himeno pmid=3096969 | #Himeno pmid=3096969 | ||
# | #Himeno_errata pmid=212400 | ||
</biblio> | </biblio> |
Revision as of 22:32, 11 July 2006
Already measured with SP1.0 (I13534.pSB1A3)
- Q04400.007 - mutated Q04400 (tetR QPI), point mutation in RBS. Works well, switches later than Q04400.
- Mnt inverters from enterobacteriophage p22
- Note: The promoter used in these inverters is missing base 13 in its 17-bp operator site [1], though I think they meant to include it (they reference the article and mention it in the registry). It's also missing a -35 site.
- Q04720 - mnt inverter, strong repressor - output not very high, and doesn't appear to be doing much (picture)
- Q04730 - mnt inverter, weak repressor - output not very high, and doesn't appear to be doing much (picture)
Measured with older versions of screening plasmid (I13537.pSB1A3, I13538.pSB1A3, or I13534.pSB4A3)
- Q04400 - Works reliably (quantitatively), switches at low input.
- Q03400 - tetR inverter, weaker RBS driving tetR. Looks like it starts to switch, but very high GFP gives very high RFP numbers. Unclear what's happening.
- Q01400 - tetR inverter, weakest RBS. Generally looks like it's stuck in the high-output stage. Some evidence suggests it might be switching, however.
- Q01530 - p22 cII inverter. Appears to be stuck in the low-output stage, perhaps due to crosstalk with CW2553/lambda.
- Q04530 - Same as Q01530, but with a stronger RBS driving the cII. Also stuck in low-output.
- Q04740 - penI [2, 3] inverter with strongest RBS. Works well, switches at higher input than Q04400.
- Q02740 - penI inverter, strong RBS. Looks like it begins to switch at high input.
- Q03740 - penI inverter, medium strength RBS. Stuck on.
To be measured/constructed/etc
- I14103 (Q04740-Q04400.007) - bistable switch one way
- I14104 (Q04400.007-Q04740) - bistable switch the other way - having trouble getting it cloned in this direction (tetR-penI) - weird stuff happening with growth rate and trying to prep the final plasmid, though colony PCR looks fine - load issues?
- Q04510 - Ligated into the screening plasmid, but somehow missed being characterized. Seems unlikely to work, but hasn't been tested.
- Q07400/Q08400/Q09400 - tetR inverters using RBS's from the Voigt lab. Not BB'd, and previous attempts were unsuccessful.
References
- Stormo GD, Strobl S, Yoshioka M, and Lee JS. Specificity of the Mnt protein. Independent effects of mutations at different positions in the operator. J Mol Biol. 1993 Feb 20;229(4):821-6. DOI:10.1006/jmbi.1993.1088 |
- Himeno T, Imanaka T, and Aiba S. Nucleotide sequence of the penicillinase repressor gene penI of Bacillus licheniformis and regulation of penP and penI by the repressor. J Bacteriol. 1986 Dec;168(3):1128-32. DOI:10.1128/jb.168.3.1128-1132.1986 |
- Snodgrass DR and Wells PW. Passive immunity in rotaviral infections. J Am Vet Med Assoc. 1978 Sep 1;173(5 Pt 2):565-8.