Team:Davidson-Missouri Western/Improvement of Pre-existing Registry Parts
From 2008.igem.org
Improved function by a modified Lac promoter - Mutations were introduced into the wild type Lac promoter that exists in the Registry. One mutant promoter, designated pLacIQ1, had improved ability to promote, as demonstrated by the following experiment.
Constructs were assembled with eiter the wild type Lac promoter or mutant promoters driving GFP expression via part number E0240. Fluorescence measurements taken at 16 or 24 hours were corrected for optical density.
16 hours
Sample | Fluorescence | Optical Density | Fluor/OD | |
---|---|---|---|---|
LB | 2 | 0.043 | 46.5 | |
E0240 | 0.67 | 1.262 | 0.53 | |
S04020 | 4.67 | 1.225 | 3.81 | |
S04021 | 21 | 1.201 | 17.49 | |
K091131 | 68 | 1.188 | 57.24 |
24 hours
Sample | Fluorescence | Optical Density | Fluor/OD | |
---|---|---|---|---|
LB | 1 | 0.047 | 21.3 | |
E0240 | 3.0 | 1.201 | 2.50 | |
S04020 | 4.0 | 1.239 | 3.23 | |
S04021 | 14.5 | 1.216 | 11.92 | |
K091131 | 71.3 | 1.207 | 59.07 |
Mutations were also introduced into the LacI repressor gene in order to improve its ability to bind to the operator site in the Lac promoter.
Part | Description | plasmid | size |
---|---|---|---|
BBa_K091121 | LacI wild-type gene | 1083 | |
BBa_K091122 | LacI_I12 | 1083 | |
BBa_K091127 | LacI_I12_X86 | 1083 |