Paulsson:Pmalkus factoids

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wish I knew

  • where is membrane added in coli?
  • how fast can you spin coli without losing cfu? stationary vs. log phase growth
  • percent of MFG-GFP fusions that are non-functional (eg in S.c. ORFeome collection)??? Partial loss of function may not be detected in viability, but rather in growth rate or protein localization?
  • coli transcriptional/proteome resources
  • model building resources

FPs

  • in vivo maturation of Venus ~7 -/+ 2.5 min (Yu and Xie, Science'06)
  • homoaffinity of purified recombinant YFP, K(d) = 0.11mM (Zacharias et al., Science v296:913-916, 2002)
  • degradation of GFP-LAA: in vivo??? in vitro <5min (Bukau, Sue Wickner)
  • degradation of N-end rule substrate ~2min (Varshavsky Science'91)
  • GFP fluorescence lifetime ~3ns (Burak says so)

coli

  • don't prewarm Amp plates at 37deg for more than ~ 30min (otherwise satellites will appear)
  • E. coli doubling times

strain W1485: 46.9' at 28deg, 24.1' at 37deg, 21.4' at 42deg (Condon and Squires, J.Bact 1995)

  • supplementing minimal media with 0.1% casamino acids attenuates GFP toxicity (Alper and Stephanopoulos, PNAS'05)
  • Compared to closed circular dsDNA, nicked circular DNA and ssDNA are 50% as efficient, linear DNA is 0.1% as efficient in transformation of E. coli.

lab strains

  • The plasmid in ST11 and ST12 (and all pPM derivatives) have an A to G mutation in the PLtet-O1 promoter (at position 1982 in pPM5). This substitution doesn't appear to alter behavior of Ptet, but I haven't done the direct comparison to original PLtet-O1.
  • PMB14 and 27 both contain integrated version of LacI AND TetR. They work equally well at repressing Plac and Ptet.
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