Difference between revisions of "Part:BBa K3081004"

 
 
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<partinfo>BBa_K3081004 short</partinfo>
 
<partinfo>BBa_K3081004 short</partinfo>
  
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Tac promoter is a hybrid between trp and lacUV5 promoters. Tac promoter is a kind of strong E. coli promoter and is IPTG inducible. pTAC promoter induces the protein dCas9,and the C terminus of dCas9 is fused to GFP coding sequence. Expression level of dCas9 induced by IPTG is tested by fluorescence of fused dCas9 linker GFP
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pTAC is a lactose-inducible promoter ,and it is also effectively induced by isopropyl-β-D-1-thiogalactoside (IPTG), which can diffuse across the cell membrane without the aid of the lactose transporter. This classic operon consists of two basic parts, a cassette for the constitutive expression of the repressor LacI and a cassette for the inducible expression of the target gene. The LacI can bind to the lac operator (lacO) region in the second cassette and trigger the repression. This repression stops transcription from the promoter itself. The derepression of transcription is carried out by inducers such as IPTG, which can remove LacI from the operator region by binding to it.
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Here, we use the pTAC promoter to drive the expression of sfGFP and we can observe the intensity of fluorescense to obtain a proper inducing concentraion.
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<center>
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https://2019.igem.org/wiki/images/6/68/T--Peking--pTAC.png
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</center>
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<center>
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Figure 1: IPTG-induced GFP expression level tested by cytometer
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</center>
  
 
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Latest revision as of 10:23, 21 October 2019


pTAC GFP

pTAC is a lactose-inducible promoter ,and it is also effectively induced by isopropyl-β-D-1-thiogalactoside (IPTG), which can diffuse across the cell membrane without the aid of the lactose transporter. This classic operon consists of two basic parts, a cassette for the constitutive expression of the repressor LacI and a cassette for the inducible expression of the target gene. The LacI can bind to the lac operator (lacO) region in the second cassette and trigger the repression. This repression stops transcription from the promoter itself. The derepression of transcription is carried out by inducers such as IPTG, which can remove LacI from the operator region by binding to it. Here, we use the pTAC promoter to drive the expression of sfGFP and we can observe the intensity of fluorescense to obtain a proper inducing concentraion.

T--Peking--pTAC.png

Figure 1: IPTG-induced GFP expression level tested by cytometer

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1472