Difference between revisions of "Part:BBa K2890001"
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A optimized frmR promoter for formaldehyde detection. | A optimized frmR promoter for formaldehyde detection. | ||
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| + | =Characterization from JNFLS2019= | ||
| + | ==1.The toxic effect of formaldehyde on the cells growth== | ||
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| + | We added the reporter gene GFP to the downstream of the promoter, then detected the tolerance of the cells and the sensitivity of the promoter to formaldehyde using BL21 as chassis. | ||
| + | Formaldehyde is toxic to cells. It can inhibit the cells growth if it is added to the culture medium too early. In order to know at which OD600 the formaldehyde should be added into the culture medium, we used 0.5mM formaldehyde to treat cells at different original OD600, then we collected different group bacteria at certain time to measure cells’ DO600 absorbance (Figure 1). The result showed that the toxic effect of formaldehyde on the cells growth was decreased with the cells original OD600 increased. | ||
| + | [[File:K2890001-1.jpg|center]] | ||
| + | ==2.The sensitivity of promoter to formaldehyde== | ||
| + | We cultured E.coli BL21 transformed with the plasmid (PfrmR-GFP) and added different concentration formaldehyde to different groups when the OD600 = 1, then we collected bacteria at certain time to measure the fluorescence and the absorbance. From the Figure 2, we found that 0.8 mM formaldehyde is the best concentration for inducing GFP expression. | ||
| + | [[File:K2890001-2.jpg|center]] | ||
| + | <br/> | ||
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Latest revision as of 03:58, 20 October 2019
optimized frmR
A optimized frmR promoter for formaldehyde detection.
Characterization from JNFLS2019
1.The toxic effect of formaldehyde on the cells growth
We added the reporter gene GFP to the downstream of the promoter, then detected the tolerance of the cells and the sensitivity of the promoter to formaldehyde using BL21 as chassis. Formaldehyde is toxic to cells. It can inhibit the cells growth if it is added to the culture medium too early. In order to know at which OD600 the formaldehyde should be added into the culture medium, we used 0.5mM formaldehyde to treat cells at different original OD600, then we collected different group bacteria at certain time to measure cells’ DO600 absorbance (Figure 1). The result showed that the toxic effect of formaldehyde on the cells growth was decreased with the cells original OD600 increased.
2.The sensitivity of promoter to formaldehyde
We cultured E.coli BL21 transformed with the plasmid (PfrmR-GFP) and added different concentration formaldehyde to different groups when the OD600 = 1, then we collected bacteria at certain time to measure the fluorescence and the absorbance. From the Figure 2, we found that 0.8 mM formaldehyde is the best concentration for inducing GFP expression.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 551