Part:BBa_K515102:Experience
Characterisation
This part (BBa K515102) has been characterised qualitatively and quantitatively for chemotactic response towards L (-) malic acid. The tests were performed across a range of different attractant concentrations to observe optimal functioning of the part. Qualitative tests were performed using semi solid agar assays, quantitative analysis was performed using high - throughput capillary assay with cell count for each capillary evaluated using FACS. Behavioral analysis of E. coli expressing this part was done using Wide - field microscopy, and manual tracking was used to evaluate tactic response. Negative control used was E. coli 5α strain with a selectable marker for ampicillin and kanamycin. As positive control chemotactic response of E. coli 5α strain to attractant L (-) serine was tested, as serine is compatible with endogenous chemotaxis system of E. coli 5α strain.
Qualitative assay
Two tests were performed to observe chemotactic response over a large range of chemoattractant concentration and to observe differences in much finer milimolar ranges of attractant concentration.
Table 1: Ranges of attractant concentration tested.
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Attractant concentration |
||||||
Test 1 |
0 M |
10-5 M |
10-4 M |
10-3 M |
10-2 M |
10-1 M |
Test 2 |
0 mM |
5 mM |
10 mM |
15 mM |
20 mM |
25 mM |
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