Team:NYMU-Taipei/ymiq3.html

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     <p><span class="subtitle">DCMU concentration and cell growth</span></p>
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     <p><span class="subtitle">Resistance of Synechococcus SP. PCC 7002 to 3 - (3,4-dichlorophenyl) - 1,1 - dimethylurea</span></p>
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     <p>This experiment is similar to the second one of Synechococcus SP. PCC 7002 testing series. The main idea was to find the suitable DCMU concentration for Synechococcus SP. PCC 7942. As a matter of fact, both wild type and sqr expressing strain are used in the experiment.  <br />
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     <p>Synechococcus sp. PCC7002 was cultured in the medium A2, rotary shaker and initial cell concentration are the same as the last experiment. DCMU is diluted with A2 medium into different concentration, and 10 mM of sodium sulfide is added to the experimental group. The measurement method and frequency of cell growth has been described previously.  <br />
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       <p><span class="subtitle">Sulfide concentration and the growth of sqr expressing strain Synechococcus SP. PCC 7942</span></p>
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       <p><span class="subtitle">Sodium sulfide concentration and cell growth</span></p>
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     <p>It was expected that  SQR expressing strain and wild type counterpart would have different growth  rate under the presence of sulfide compounds. Though sulfide is naturally toxic  to <em>Synechococcus SP. PCC 7942</em>, the  strain with sqr should be able to metabolize sulfide and therefore prosper. As  the result, we analyze H2S amount to detect whether sqr gene work or not. Therefore, we perform Chemical microvolume  turbidimetry method to detect H2S concentration (see <strong>Sulfur Oxide Terminator part</strong>)<br />
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     <p>With the result of previous experiment, the concentration of both sodium sulfide and DCMU is adjusted. Sodium sulfide is diluted from 40mM to 2.5mM, while DCMU 0.5 μM is added into the experimental group.<br />
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Revision as of 19:59, 26 October 2012

NYMU iGEM

Experiments

Resistance of Cyanobacteria (Synechococcus SP. PCC 7002) to Sulfide compound

Synechococcus sp. PCC7002 was cultured in the medium A2, a modification of Stevens et al. (1973) on a rotary shaker (100 rpm). The initial concentration of cells is controlled to an OD730 of 0.1 in fresh medium. Different concentration of sodium sulfide is added, and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) was put into experimental group. Cells are grown in the six-well-plates and monitored under OD730 every 24 hour.

Resistance of Synechococcus SP. PCC 7002 to 3 - (3,4-dichlorophenyl) - 1,1 - dimethylurea

Synechococcus sp. PCC7002 was cultured in the medium A2, rotary shaker and initial cell concentration are the same as the last experiment. DCMU is diluted with A2 medium into different concentration, and 10 mM of sodium sulfide is added to the experimental group. The measurement method and frequency of cell growth has been described previously.

Sodium sulfide concentration and cell growth

With the result of previous experiment, the concentration of both sodium sulfide and DCMU is adjusted. Sodium sulfide is diluted from 40mM to 2.5mM, while DCMU 0.5 μM is added into the experimental group.

Sulfide oxidation in Escherichia coli expressing sulfide-quinone reductase gene

Repots have it that Escherichia coli can express functional sulfide-quinone reductase (SQR). Therefore, we slightly adjusted the previous experiment and applied to the SQR gene from Synechococcus SP. PCC 7002. With methylene blue method, we would test the efficiency of SQR sulfide oxidation. Since such method involved in measurement of optical density, it is more appropriate to perform such experiment on colorless bacteria instead of engineered cyanobacteria strain.

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