Difference between revisions of "Part:BBa K2967019"

Revision as of 06:51, 10 October 2019

The yeaR promoter will initiate the express of IL-10 when nitric oxide appear

In order to see whether the nitric oxide sensor will work when it comes to expressing a certain protein, we add the IL-10 downstream and demonstrated the western blotting. It has been proved that the expression level can be induced when adding nitric oxide.

Figure 1. Diagram of expressing vector in backbone pCDFDuet-1. Promoter yeaR is an nitric oxide inducible promoter. Gene hIL-10 is one of our target protein.

Figure 2. Expressing vector is transformed into BL21 strain. After induction of nitric oxide, the culture is incubated at 37℃ for 2 hours. Western blotting demonstrats the expression level of hIL-10. In order to equate the total number of cells in each lane, dilution is made to keep each sample at the same OD and sample volume.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 https://static.igem.org/mediawiki/parts/thumb/3/3c/T--NEU_China--part--pyear%2Bli10-2.png/800px-T--NEU_China--part--pyear%2Bli10-2.png
-'''Figure 2. Expressing vector is transformed into BL21 strain.''' After induction of nitric oxide, the culture is incubated at 37℃ for 2 hours. Western blotting demonstrats the expression level of hIL-10. In order to equate the total number of cells in each lane, dilution is made to keep each sample at the same OD and μL of each sample is uploaded to each well.
+'''Figure 2. Expressing vector is transformed into BL21 strain.''' After induction of nitric oxide, the culture is incubated at 37℃ for 2 hours. Western blotting demonstrats the expression level of hIL-10. In order to equate the total number of cells in each lane, dilution is made to keep each sample at the same OD and sample volume.
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