Difference between revisions of "Part:BBa K3633016"
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==Description== | ==Description== | ||
| − | This is a common promoter that can be induced by IPTG. The iGEM20_Shanghai_SFLS_SPBS | + | This is a common promoter that can be induced by IPTG. The iGEM20_Shanghai_SFLS_SPBS used the promoter to accomplish experiments, including the production of melanin and the test of IPTG-induced sfGFP production. |
==Experiment and Results== | ==Experiment and Results== | ||
===Successful production of melanin in E.coli BL21(DE3) and vibrio natriegens with promoter pTac=== | ===Successful production of melanin in E.coli BL21(DE3) and vibrio natriegens with promoter pTac=== | ||
| − | We constructed the plasmid pTac-Tyr1 and successfully transformed the plasmid into E. coli BL21(DE3) and Vibrio natriegens ATCC 14048. We added 2.5 mg L-DOPA | + | We constructed the plasmid pTac-Tyr1 and successfully transformed the plasmid into E. coli BL21(DE3) and Vibrio natriegens ATCC 14048. We added 2.5 mg L-DOPA and 0.4 mg L-Tyrosine as the substrates and 1 mM IPTG as induction. We cultured the bacteria in shakers (220 rpm) at 37℃. The result shows that melanin was produced. Although functioning, the promoter's effect on this specific enzyme was not as good as the constitutive promoter J23102. |
| − | [[File:T--Shanghai_SFLS_SPBS--Melanin Result 7.png|600px|center|thumb|Fig 3.Production of melanin in E. coli BL21(DE3) and Vibrio natriegens at 37℃ in 78 h. The horizontal axis is time (hours), and the vertical axis is absorbance of the bacterial solution at 400 nm. ]] | + | [[File:T--Shanghai_SFLS_SPBS--Melanin Result 7.png|600px|center|thumb|Fig 3.Production of melanin in E. coli BL21(DE3) and Vibrio natriegens at 37℃ in 78 h. The horizontal axis is time (hours), and the vertical axis is the absorbance of the bacterial solution at 400 nm. ]] |
==Sequence & Features== | ==Sequence & Features== | ||
Revision as of 04:30, 27 October 2020
pTac promoter
Description
This is a common promoter that can be induced by IPTG. The iGEM20_Shanghai_SFLS_SPBS used the promoter to accomplish experiments, including the production of melanin and the test of IPTG-induced sfGFP production.
Experiment and Results
Successful production of melanin in E.coli BL21(DE3) and vibrio natriegens with promoter pTac
We constructed the plasmid pTac-Tyr1 and successfully transformed the plasmid into E. coli BL21(DE3) and Vibrio natriegens ATCC 14048. We added 2.5 mg L-DOPA and 0.4 mg L-Tyrosine as the substrates and 1 mM IPTG as induction. We cultured the bacteria in shakers (220 rpm) at 37℃. The result shows that melanin was produced. Although functioning, the promoter's effect on this specific enzyme was not as good as the constitutive promoter J23102.
Sequence & Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]