Difference between revisions of "Part:BBa K3081004"
 
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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.
 
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 dCas9 and we can observe the fluorescense of the GFP fused to C-terminal of dCas9 to obtain a proper inducing concentraion.
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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