Difference between revisions of "Part:BBa K733018:Experience"

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<p>Compared with the results of the efficiency of the xylose inducible promoter detected by the HKT, our consequence differed from them in some aspects, or better. We had used the FCM to detect it. There was an obvious tendency that the efficiency of the promoter increased as the xylose concentration gathered up. What was amazing was that the fluorescence we got was 10 times brighter than the result the HKT got, which appeared to be great.</p>
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<p>But when the concentration of the xylose was 0.03% and 0.3%, we got an unexpected result that the expression of the GFP fell after rise. What’s more. While the concentration of the xylose was 10%, the expression of the GFP decreased, which corresponded with the result of the HKT.</p>
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<I>[http://2013.igem.org/Team:SUSTC-Shenzhen-A 2013SUSTC-Shenzhen-A]</I>
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<p>First we used the microplate to detect the fluorescence of this part, because we could not see the transition point and we were wondering what it would be look like. The data we got is not so good, for we didn’t see the transition point obviously, and also has some differences comparing to the data from HKUST. According to the detecting from HKUST, fluorescence produced in about 5% xylose is more than that in 10% xylose, however what we got is fluorescence produced in 5% xylose than that in 10% xylose.</p>
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<p>Since we didn’t get what we really want using microplate, we tried FCM this time. In 0% xylose culture, we almost got no fluorescence produced and in 0.003% xylose culture we saw obvious fluorescence came out. There was an obvious tendency that the efficiency of the promoter increased as the xylose concentration gathered up. However, with the increase of xylose concentration, the fluorescence increased choppily. What was amazing was that the fluorescence we got was 10 times brighter than the result the HKT got, which appeared to be great.</p>
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<p>Another interesting thing is most of the cell we detected had no fluorescence produced. Then we guess that may because the xylose molecules taken in the different cell are different, or may because the copy number in each cell is different. Maybe it has some relationship with epigenetics. </p>
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Latest revision as of 19:50, 27 September 2013


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[http://2013.igem.org/Team:SUSTC-Shenzhen-A 2013SUSTC-Shenzhen-A]

Xylose-gfp.jpg

First we used the microplate to detect the fluorescence of this part, because we could not see the transition point and we were wondering what it would be look like. The data we got is not so good, for we didn’t see the transition point obviously, and also has some differences comparing to the data from HKUST. According to the detecting from HKUST, fluorescence produced in about 5% xylose is more than that in 10% xylose, however what we got is fluorescence produced in 5% xylose than that in 10% xylose.

xylose

Since we didn’t get what we really want using microplate, we tried FCM this time. In 0% xylose culture, we almost got no fluorescence produced and in 0.003% xylose culture we saw obvious fluorescence came out. There was an obvious tendency that the efficiency of the promoter increased as the xylose concentration gathered up. However, with the increase of xylose concentration, the fluorescence increased choppily. What was amazing was that the fluorescence we got was 10 times brighter than the result the HKT got, which appeared to be great.

Another interesting thing is most of the cell we detected had no fluorescence produced. Then we guess that may because the xylose molecules taken in the different cell are different, or may because the copy number in each cell is different. Maybe it has some relationship with epigenetics.


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