Part:BBa_K3317051
NhaS with an RFP reporter
It produces a protein called NhaS BBa_K1255000 that is described as a sodium ion binding protein in Bacillus pseudofirmus . The RFP sequence has been added to inform its transcription with a red fluorescence indicator. This part was characterized in Escherichia coli to generate a sodium ion capture system from the medium in which it is found. Because this piece is optimized with a constitutive promoter, its performance was evaluated by its ability to capture sodium ions. Krulwich in 1994 mentions that NhaS could act as a channel of ions in the membrane that promotes the mechanism of antiportation (Image 1), introducing sodium ions so that the cell finds an equality between the osmotic pressures both inside and in the exterior. In this way the cell would be able to avoid its lysis by a difference in the salt concentrations present both inside and outside it.
In 2014, the [http://2014hs.igem.org/Team:CIDEB-UANL_Mexico/labwork_results CIDEB-UANL_Mexico] high school synthesized the nhaS gene and introduced it into E. coli cells to test its resistance to saline environments. It was found that the transformed cells are capable of withstanding high concentrations of salinity in the medium in which they are found, since they managed to have a growth in media at concentrations of up to 15% NaCl. This resistance to saline environments and its hypothetical capacity of ion absorption given by the NhaS protein, will be used in this work as part of a biological desalination process together with a protein capable of fixing itself to silica surfaces, thus facilitating the recovery of cells that previously trapped sodium ions due to the expression of the NhaS protein and employ them as well as an organism with high resistant to salty media.
The function of the NhaS protein was evaluated by comparing salt consumption between the recombinant strain and a control without the gene, using electro conductivity as an indirect indicator of sodium ions dissolved in the culture medium. The statistical analysis of the results produced by the experiment showed statistically significant differences with a p-value of 0.008 below the 0.05 alpha between the control and the samples. Since the difference between these groups is the presence of the nhas gene, from this you can consider this factor as the cause of increased consumption in the middle salt. Given the results, it is plausible to state that the nhaS gene allows greater absorption of sodium ions compared to those that lack it.
Characterization Experiments and Results
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 849
Illegal AgeI site found at 961 - 1000COMPATIBLE WITH RFC[1000]
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