Team:Hamburg/Lab Results
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Lab Results
Biochemistry
In this project we were able to produce several parts, but unfortunately we only had enough time to charcterize some of them. To these belong promotor + tired + terminator and promtor + sfp, which were charcterized via kinetic measurements. The measured OD values before and after induction with 400 mM IPTG (+ 200 µg cystein) are shown in tables 1 and 2.
Table 1: OD values before induction| BL21 | tired | sfp | |
|---|---|---|---|
| t = 0 | 0.1250 | 0.0364 | 0.0260 |
| t = 70 | 0.3473 | 0.0644 | 0.1154 |
| t = 120 | 1.0422 | 0.3031 | 0.4058 |
| t = 180 | 1.5018 | 0.4356 | 0.9930 |
Chemistry
The products pyochelin, NH-nor-pyochelin and 4,5-Dihydro-2-(2,4-dihydroxyphenyl)thiazole-4(R)-carboxylic acid were synthetized.
The presence and purity of the products i>NH-nor-pyochelin and 4,5-Dihydro-2-(2,4-dihydroxyphenyl)thiazole-4(R)-carboxylic acid was confirmed with 1H-, 13C- and 2D NMR spectroscopy while the presence of pyochelin was confirmed using only 1H-NMR, since 2D spectra were not available at that point. The pyochelin was contaimnated to a certain degree, but it was nevertheless used for subsequent testings. The amount of pyochelin was not sufficient for purification.
UV-Vis spectra of Pyochelin, NH-nor-pyochelin and 4,5-Dihydro-2-(2,4-dihydroxyphenyl)thiazole-4(R)-carboxylic acid were then recorded. To investigate complex formation with iron(III) and gallium (III), solutions of salts including both metals (gallium(III) nitrate and iron(III) chloride) were mixed with the siderophore solutions. UV-Vis spectra were recorded to investigate the kinetics of the complex formations (see here). Finally, siderophore-gallium complexes were created and used for toxicity tests.
Nanoscience
Sadly our first charge of microfluidic devices broke down when we tried to coat them with an ECM-solution to grow our cells on. And we were not able to produce a second charge before wiki freeze.
