Team:USP-UNESP-Brazil/Associative Memory/Experiments

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(Tests of System "Cin" of quorum sensing)
(Test for Internal feedback and basal production rate)
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In the case of satisfactory results in the previous tests, this one will probably work as well. The main objective is to estimate if the rate of basal expression is not excessive, in other words, if the positive feedback system is low enough. For this, we can compare the rate of basal fluorescence using the graph made on “HSL  production test”  as parameter.
In the case of satisfactory results in the previous tests, this one will probably work as well. The main objective is to estimate if the rate of basal expression is not excessive, in other words, if the positive feedback system is low enough. For this, we can compare the rate of basal fluorescence using the graph made on “HSL  production test”  as parameter.
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If the fluorescence doesn’t  appear, there are 2 main possibilities:  The feedback system isn’t working or it’s too low. In order to verify this issue, the system could be stimulated by insertion of QS substance, measuring the period of glowing and comparing it to the glowing time of bacteria from lineage 2 (which doesn’t have feedback system).
If the fluorescence doesn’t  appear, there are 2 main possibilities:  The feedback system isn’t working or it’s too low. In order to verify this issue, the system could be stimulated by insertion of QS substance, measuring the period of glowing and comparing it to the glowing time of bacteria from lineage 2 (which doesn’t have feedback system).
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It is possible to verify how much HSL will be necessary to activate the QS of the system. In order to do this, will be necessary to take samples from the growth medium from lineage 1 with several different “IPTG inductions”. It is likely to exists a threshold concentration of the QS substance in which the QS starts and the fluorescence of the system keeps glowing without external influence.
It is possible to verify how much HSL will be necessary to activate the QS of the system. In order to do this, will be necessary to take samples from the growth medium from lineage 1 with several different “IPTG inductions”. It is likely to exists a threshold concentration of the QS substance in which the QS starts and the fluorescence of the system keeps glowing without external influence.
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Tests  of Rhl system of Quorum Sensing.
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===Tests  of Rhl system of Quorum Sensing.===
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Just as previously, we intend to check the operation of the QS system, but this time with the rhl system. We will check:  production of rhIL enzyme, levels of activation and repression of rhlR substance and ,lastly, intensity positive feedback system, comparing levels of activation and inhibition with tested previously.
Just as previously, we intend to check the operation of the QS system, but this time with the rhl system. We will check:  production of rhIL enzyme, levels of activation and repression of rhlR substance and ,lastly, intensity positive feedback system, comparing levels of activation and inhibition with tested previously.
We will do the constructions presented in Figure 8 in addition to tests similar to the system in each Cin.
We will do the constructions presented in Figure 8 in addition to tests similar to the system in each Cin.
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[[File:0014.jpg|center|600px|caption|]]
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"Figure 8"
===BHL production test.===
===BHL production test.===

Revision as of 21:23, 22 September 2012