Team:NCKU Tainan/Contribution

Contribution

Introduction


Nitrate is toxic to fish or shrimp in water. When nitrate reaches a certain concentration level, it will be a harsh condition for aquatic animal. In order to maintain good quality of water, concentration of nitrate should be taken more seriously. Previously, BCCS Bristol 2010 have constructed BBa_K381001, a nitrate sensing GFP composite. We want to take advantage of it and refine the characterization of nitrate sensing. Especially to fit aquaculture condition, we focus on the range of 0 to 100 ppm.

Description


Part: BBa_K381001 was first designed by Katharine Coyte from team BCCS-Bristol in iGEM 2010. It is a nitrate reporter, PyeaR - GFP composite. The team BCCS-Bristol only tested the biobrick’s sensitivity at a higher concentration, but the Nitrate level of aquatic water usually won’t reach 1 mM. Therefore, we carry out experiment by testing the fluorescence intensity with ppm level of potassium nitrate, which is much lower than all the teams before. The results are shown below.

contribution image 1

Figure 1. The fluorescence intensity of different nitrate concentration taken the logarithm at 8, 12 and 24 hours.

It shows that the biobrick BBa_K381001 has nice sensitivity not just in high nitrate concentration, but low as well. Many teams had done improvements of K381001, but our team brings out the most beautiful data. And we also take X axis logarithm, which can make our data become linear. Not only easier to read, but also more meaningful.

The data are shown as below.

ppm conc 8 hours 12 hours 24 hours
0 454.6 546.8 620.4
2 689.9 754.0 821.1
4 766.8 866.8 922.8
6 816.4 916.1 972.6
8 831.8 931.4 985.1
10 900.1 998.9 1058.8
20 962.4 1093.0 1148.2
40 1090.9 1200.9 1252.1
60 1129.6 1253.0 1295.0
80 1240.3 1341.7 1373.6
100 1259.7 1385.9 1449.9