Team:Glasgow/Notebook
James’ muffins
Talked to Professors Marshall Stark and John Christie about our project.
Repressors
Ordered PPhlF and PSrpR with BioBrick prefix and suffix as oligos. Ordered PhlF and SrpR as G-blocks, with ribosome binding site B0032, terminator B0010+ (a longer version of B0010, that we will characterise and submit as a new BioBrick) and BioBrick prefix and suffix. Got E5501 and I13500 – GFP (E0040) with ribosome binding sites B0032 and B0034 respectively – in pSB1C3 in DH5α from glycerol stocks from last year’s iGEM team.
UVA sensor
Designed primers to PCR UirS and UirR from Synechocystis PCC6803 genomic DNA with ribosome binding site B0032 and BioBrick prefix and suffix. Tested UVB sensitivity of strains TOP10 and DH5α (recA- - very poor DNA repair), DS941 (recF- - limited DNA repair), and MG1655.Z1 (wild type for DNA repair).
Bioluminescence
pBAD.LuxCDABEG (K325909) and pBAD.LuxCDABEG.LumP (K769020) off distribution plates and transformed into TOP10 and DH5α. Designed primers to PCR LuxA, B, C, D, E, and G individually with ribosome binding site B0032 and BioBrick prefix and suffix from K325909.
Vilija’s carrot cake
So good, honestly.
