Team:SDU-Denmark/K343007

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(Experiment 2)
(K343007)
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<div id="parts">
<div id="parts">
== K343007 ==
== K343007 ==
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The part K343007 (from now on shortly called PS) is a generator for the SopII-HtrII photosensor from ''Natronomonas pharaonis'' coupled to ''E. Coli's'' chemotaxis pathway via the ''Salmonella enterica'' protein Tar. This part's effect on the system is to make ''E. Coli'' phototactic, so that it becomes aware of different light conditions. So for characterization of this part we made examination of motility and motility patterns our first priority and plasmid stability and growth of the cells our second. <br>
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The part K343007 (hereafter referred to as "PS") is a generator for the SopII-HtrII photosensor from ''Natronomonas pharaonis'' coupled to ''E. coli's'' chemotaxis pathway via the ''Salmonella enterica'' protein Tar. This part's effect on the system is to make ''E. coli'' phototactic, so that it becomes aware of different light conditions. So for characterization of this part we made examination of motility and motility patterns our first priority and plasmid stability and growth of the cells our second. <br>
There is a wide range of motility assays for chemotaxis in bacteria, this meant that we had a broad spectrum of experiments to choose from, which just had to be tweaked for making them suited for the analysis of phototaxis. The two experiments we chose for analysing the effect of this part (PS), were growth of the bacterial cultures in semi-solid agar and computer analysis of swimming motility through video microscopy.<br>
There is a wide range of motility assays for chemotaxis in bacteria, this meant that we had a broad spectrum of experiments to choose from, which just had to be tweaked for making them suited for the analysis of phototaxis. The two experiments we chose for analysing the effect of this part (PS), were growth of the bacterial cultures in semi-solid agar and computer analysis of swimming motility through video microscopy.<br>
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[[Image:Team SDU-Denmark OD WT+PS.JPG|thumb|center|400px|Growth curve showing measured OD at 550nm. Samples were taken every hour over a 12 hour perion + one sample after 24 hours. In graph ''E. coli'' strain MG1655-pSB3T5-k343007 and MG1655-pSB1C3-K343007 is compaired with Wild type ''E. coli'' strain MG1655 showing no significant difference between the three graphs. No Lag phase is seen. All data can be seen under [https://static.igem.org/mediawiki/2010/2/21/Team_SDU_Denmark_Growth_rate_assay_2_PS.zip Raw data] ]]<br><br>
[[Image:Team SDU-Denmark OD WT+PS.JPG|thumb|center|400px|Growth curve showing measured OD at 550nm. Samples were taken every hour over a 12 hour perion + one sample after 24 hours. In graph ''E. coli'' strain MG1655-pSB3T5-k343007 and MG1655-pSB1C3-K343007 is compaired with Wild type ''E. coli'' strain MG1655 showing no significant difference between the three graphs. No Lag phase is seen. All data can be seen under [https://static.igem.org/mediawiki/2010/2/21/Team_SDU_Denmark_Growth_rate_assay_2_PS.zip Raw data] ]]<br><br>
From our data we see no significant difference between the plasmid carrying bacteria and the wild type. This can be said to be quite conteradictory to our results obtained from the stability assay. The transitory stability of pSB1C3-K343007 suggests that it is highly unfavorable for the bacteria, wherefore it might be expected that the growth of the bacteria containg this plasmid would be affected. Thus, however much a disadvantage the plasmid pose to the bacteria, their growth are not significantly influenced by the plasmid. The added reproduction load due to the plasmids, might also prolong the lag phase of the bacteria. Whether this is the case can not be concluded based on this experiment as no lag phase was observed in this experiment.<br><br>
From our data we see no significant difference between the plasmid carrying bacteria and the wild type. This can be said to be quite conteradictory to our results obtained from the stability assay. The transitory stability of pSB1C3-K343007 suggests that it is highly unfavorable for the bacteria, wherefore it might be expected that the growth of the bacteria containg this plasmid would be affected. Thus, however much a disadvantage the plasmid pose to the bacteria, their growth are not significantly influenced by the plasmid. The added reproduction load due to the plasmids, might also prolong the lag phase of the bacteria. Whether this is the case can not be concluded based on this experiment as no lag phase was observed in this experiment.<br><br>
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= References =
= References =

Revision as of 00:32, 28 October 2010