Paris/July 3
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Revision as of 10:05, 11 July 2007
Contents |
Preparation of the P1 stock on the w121 strain.
See protocols. This preparation is necessary for transduction of DapA deletion in MG1655 strain.
Kinetic measurements:
As previously described, we want to measure :
- Growth of DapA- strain (w121) relative to the concentration of DAP in the medium :
- The excretion of Dap by MG1655 DapA+ strain by an indirect way
Growth of DapA- strain (w121) relative to the concentration of DAP in the medium
We measure the kinetic of the w121 as a function of DAP concentration
[DAP] = 5mM
Kinetic Array :Growthing of w121 strain as a function of DAP concentration (1µL w121 strain incubated ON) | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
A | {{{A11}}} | {{{A12}}} | ||||||||||
B | 200µL LB + 0µL DAP | 200µL LB + 2µL DAP | 200µL LB + 4µL DAP | 200µL LB + 8µL DAP | 200µL LB + 12µL DAP | {{{B11}}} | {{{B12}}} | |||||
C | {{{C11}}} | {{{C12}}} | ||||||||||
D | {{{D11}}} | {{{D12}}} | ||||||||||
E | {{{E11}}} | {{{E12}}} | ||||||||||
F | {{{F11}}} | {{{F12}}} | ||||||||||
G | {{{G11}}} | {{{G12}}} | ||||||||||
H | {{{H1}}} | {{{H2}}} | {{{H3}}} | {{{H4}}} | {{{H5}}} | {{{H6}}} | {{{H7}}} | {{{H8}}} | {{{H9}}} | {{{H10}}} | {{{H11}}} | {{{H12}}} |
See Results.
Measuring excretion of Dap by MG1655 DapA+ strain in an indirect manner
We measure the growth of DapA- strain in the filtered growth medium of previously incubated MG1655 strain (S0).
S0 = Centrifuged and filtered medium of MG1655 ON
[DAP] = 5mM
Kinetic Array :Growthing of w121 strain 'supplemented' with S0 (1µL w121 strain incubated ON) | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
A | {{{A11}}} | {{{A12}}} | ||||||||||
B | 200µL S0 + 0µL DAP | 200µL S0 + 2µL DAP | 200µL S0 + 4µL DAP | 200µL S0 + 8µL DAP | 200µL S0 + 12µL DAP | {{{B11}}} | {{{B12}}} | |||||
C | {{{C11}}} | {{{C12}}} | ||||||||||
D | {{{D11}}} | {{{D12}}} | ||||||||||
E | {{{E11}}} | {{{E12}}} | ||||||||||
F | {{{F11}}} | {{{F12}}} | ||||||||||
G | {{{G11}}} | {{{G12}}} | ||||||||||
H | {{{H1}}} | {{{H2}}} | {{{H3}}} | {{{H4}}} | {{{H5}}} | {{{H6}}} | {{{H7}}} | {{{H8}}} | {{{H9}}} | {{{H10}}} | {{{H11}}} | {{{H12}}} |
Acinetobacter calcoaceticus ADP1 culture
We received the strain Acinetobacter calcoaceticus ADP1 from Pasteur Institute (Center of biological resources of the Pasteur Institute): code CIP 104273 (ATCC number 33305). The strain was dehydrated and cultured following this protocol on LB agar plate at 30°C overnight: