Difference between revisions of "Part:BBa K3633016"
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==Description== | ==Description== | ||
| − | This is a common promoter that can be induced by IPTG. | + | This is a common promoter that can be induced by IPTG. iGEM20_Shanghai_SFLS_SPBS used this promoter to express tyrosinase (Tyr1) and produce melanin. |
==Experiment and Results== | ==Experiment and Results== | ||
| − | ===Successful production of melanin in E.coli BL21(DE3) | + | ===Successful production of melanin in E.coli BL21(DE3) 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. | 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. | ||
Latest revision as of 10:08, 27 October 2020
pTac promoter
Description
This is a common promoter that can be induced by IPTG. iGEM20_Shanghai_SFLS_SPBS used this promoter to express tyrosinase (Tyr1) and produce melanin.
Experiment and Results
Successful production of melanin in E.coli BL21(DE3) 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]