Difference between revisions of "Part:BBa K1420006"
(→Experimental Results) |
(→Experimental Results) |
||
Line 7: | Line 7: | ||
== Experimental Results == | == Experimental Results == | ||
− | <p>In order to measure the ''phsABC'' | + | <p>In order to measure the thiosulfate reducing activity of ''phsABC'' of NaS<sub>2</sub>O<sub>3</sub> to H<sub>2</sub>S, the gene was first inserted into the pBBRBB vector with a constitutive P<sub>lac</sub> promoter and transformed into K12 strain ''E. coli'' cells. As a control, pBBRBB::GFP was tested under the same conditions. The pBBRBB:phsABC K12 and pBBRBB::GFP K12 cells were grown in three test tubes each containing heavy metal tryptone medium as well as 3mM NaS<sub>2</sub>O<sub>3</sub>. A third set of test tubes were set up with the same contents except without cells as an additional negative control. After 24 hours, the tubes appeared as displayed in ''Figure 3''. The K12 cells contained The exact amount of H<sub>2</sub>S present in each of three different sets of tubes was then measured using a hydrogen sulfide assay (''Figure 4''). The numerical results are displayed in ''Figure 5'' </p> |
<p></p> | <p></p> | ||
[[File:experiment_1.jpg|950px|center|alt text]] | [[File:experiment_1.jpg|950px|center|alt text]] | ||
''Figure 3.'' | ''Figure 3.'' | ||
+ | <p></p> | ||
+ | [[File:assay.jpg|950px|center|alt text]] | ||
+ | ''Figure 4.'' 1:10 dilution of cell samples in Cline's Reagent used for hydrogen sulfide assays. | ||
<p></p> | <p></p> | ||
Revision as of 00:39, 17 October 2014
phsABC Generator
Summary
The (phsABC) gene from Salmonella enterica serovar Typhimurium LT2 encodes thiosulfate reductase, which catalyzes the stoichiometric production of hydrogen sulfide and sulfite from thiosulfate for heavy metal removal by precipitation.
Experimental Results
In order to measure the thiosulfate reducing activity of phsABC of NaS2O3 to H2S, the gene was first inserted into the pBBRBB vector with a constitutive Plac promoter and transformed into K12 strain E. coli cells. As a control, pBBRBB::GFP was tested under the same conditions. The pBBRBB:phsABC K12 and pBBRBB::GFP K12 cells were grown in three test tubes each containing heavy metal tryptone medium as well as 3mM NaS2O3. A third set of test tubes were set up with the same contents except without cells as an additional negative control. After 24 hours, the tubes appeared as displayed in Figure 3. The K12 cells contained The exact amount of H2S present in each of three different sets of tubes was then measured using a hydrogen sulfide assay (Figure 4). The numerical results are displayed in Figure 5
Figure 3.
Figure 4. 1:10 dilution of cell samples in Cline's Reagent used for hydrogen sulfide assays.
References
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 135
Illegal BglII site found at 2197
Illegal BamHI site found at 141 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 249
Illegal AgeI site found at 2772
Illegal AgeI site found at 3555 - 1000COMPATIBLE WITH RFC[1000]