IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-6-29

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Contents

Agar plate experiment

Yesterday we placed a clean agar plate in our incubator, with the lid slightly off-center, exposing the inside. This morning it was almost completely dried up. Now we know it's possible for an agar plate to dry up overnight under incubator conditions, which explains how our PCC 7942 plate died between last Thursday and last Friday.

Primer design

Peng h4xed like a madman to design our first primers, which we will use to PCR extract the entire KaiABC region from the PCC 7942 chromosome, and prepare it for BioBricking.

Long term project outline

See here.

Sigma factors

Roles for Sigma Factors in Global Circadian Regulation of the Cyanobacterial Genome-- Usha Nair,, Jayna L. Ditty, Hongtao Min, and Susan S. Golden. Useful?

Choosing plasmids

Prof. Golden has kindly offered to ship us plasmids from her lab. We need to decide what we're looking for. All plasmids will be delivery vehicles for cyanobacteria. That is, they will contain Neutral Site 1 (NS1) or Neutral Site 2 (NS2) homologous regions for homologous recombination with the cyanobacteria genome.

We will want...

  1. Generic plasmid to confer antibiotic resistance, which we will H.R. with PCC 7942. This will reduce the risk of contamination and make labwork easier.
  2. Plasmid with:
    • Different antibiotic resistance from the first
    • luxAB and luxCDE genes with a circadian-dependent promoter in front, e.g. PpsbaAI::luxAB and PpsbAI::luxCDE
    • pAM2195 fits this profile
  3. Same as above but with luc instead of lux?
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