Difference between revisions of "Part:BBa K3633016"

 
 
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<partinfo>BBa_K3633016 short</partinfo>
 
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pTac promoter
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==Description==
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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.
  
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==Experiment and Results==
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===Usage and Biology===
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===Successful production of melanin in E.coli BL21(DE3) with promoter pTac===
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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.
  
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[[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. ]]
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<span class='h3bb'>Sequence and Features</span>
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==Sequence & Features==
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<span class='h3bb'></span>
 
<partinfo>BBa_K3633016 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3633016 SequenceAndFeatures</partinfo>
  

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.

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


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]