Difference between revisions of "Part:BBa K518002"

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タグ<div></div>はある一定の領域を区切る役割があります。styleプロパティでfloat:left（左に浮く）とすると、次の要素が右に回り込みます。ほっとくと全部まわりこむので、回り込んでほしくないところでdiv style="clear:both"（両側に解除）とした面積０のカラの要素で回り込みを解除してます。
  
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応用で図を右（左でもよい）に表示して、続く文章を左に回り込ませたりな表示もできます。一応、適当なところでclearで忘れずに解除しておいてください。スタイルが崩れるのを防止できます。
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応用で図を右（左でもよい）に表示して、続く文章を左に回り込ませたりな表示もできます（下参照）。一応、適当なところでclearで忘れずに解除しておいてください。スタイルが崩れるのを防止できます。
 
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<Fig.2: Time-dependent luminescence measurement of renilla luciferase.>
 
<Fig.2: Time-dependent luminescence measurement of renilla luciferase.>
  
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[[Image:Promoter_Analysis.png |550px]]
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<Fig.3: Demonstration of promoter evaluation. Relative Promoter Unit (RPU) is calculated as: (maximum of firefly luminescence) / (maximum of renilla luminescence).
 
<Fig.3: Demonstration of promoter evaluation. Relative Promoter Unit (RPU) is calculated as: (maximum of firefly luminescence) / (maximum of renilla luminescence).
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Revision as of 09:14, 30 September 2011

Firefly-renilla dual luciferase assay kit

Luciferase assay is one of the most popular reporter assay system for quantitatively measuring the function of promoters and other cis-elements. The wide-ranging and quantitative detection is the prominent feature of this assay; light emmision from 10~(-18) ― 10~(-11) mol protein can be quantitatively measured using luminometer (see our results). A promoter or other cis-element to be analysed can be ligated above this part. Its effect will be measured as an altered firefly luciferase expression. Meanwhile, renilla luciferase expression can be used to as an internal control.

Calibration

For detailed data, see [http://2011.igem.org/Team:UT-Tokyo our page] and BBa_K518000.

Usage

We performed a test of this cassette using BBa_J23119 and BBa_R0011. We successfully verified the IPTG-dependence of BBa_R0011, and at the same time, the potency of this part as an evaluation tool. For experimental and analytical details, see [http://2011.igem.org/Team:UT-Tokyo our page].

Firefly1.png

Firefly2.png

<Fig.1: Time-dependent luminescence measurement of firefly luciferase. The first graph shows the raw data obtained by triplicate experiments. Data is presented as mean ± S.D.. The second graph is a model fitting according to the enzymatic kinetics theory.>

Renilla1.png

Renilla2.png

<Fig.2: Time-dependent luminescence measurement of renilla luciferase.>

Promoter Analysis.png

<Fig.3: Demonstration of promoter evaluation. Relative Promoter Unit (RPU) is calculated as: (maximum of firefly luminescence) / (maximum of renilla luminescence).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1790
    Illegal NheI site found at 1813
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2392
    Illegal SapI.rc site found at 827