Difference between revisions of "Part:BBa K2865006"
 
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<partinfo>BBa_K2865006 short</partinfo>
 
<partinfo>BBa_K2865006 short</partinfo>
 
<p class="MsoNormal"><span lang="EN-US">
 
<p class="MsoNormal"><span lang="EN-US">
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This part contains CMV promoter, eGFP reporter in RFC25 format(BBa_K2865005), and SV40 polyA terminator (BBa_K2865004). This part is constructed as a control with our part BNP promoter-eGFP-poly(A) (BBa_K2865008). </span></p>
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This part contains CMV promoter, eGFP reporter in RFC25 format(<partinfo>BBa_K2865005</partinfo>), and SV40 polyA terminator (<partinfo>BBa_K2865004</partinfo>). This part is constructed as a control with our part BNP promoter-eGFP-poly(A) (<partinfo>BBa_K2865008</partinfo>). </span></p>
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<p class="MsoNormal" style="text-indent: 0cm;"><span lang="EN-US">&nbsp;</span></p> [[File:T--SMMU-China--cep tushi.png|500px|thumb|center|Fig.1 the construction of part]]<!-- Add more about the biology of this part here-->
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<!-- Add more about the biology of this part here
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===Experiment===
 
===Experiment===
 
<B>Fluorescence microscope</B>
 
<B>Fluorescence microscope</B>
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<p class="MsoNormal"><span lang="EN-US">We transfected H9C2 cell with target plasmids, BBa_K2865006 and BBa_K2865005, via liposome transfection. After 48 hours’ expression, we detected the cells using fluorescence microscope. And we have taken clearly photos. Wo all know that the CMV promoter is widely and commonly used in plasmids or viruses designed for gene expression in mammalian cells since it is a very strong promoter and drives constitutive expression of genes under its control. And we have to admit that the transfection efficiency is low. </span></p>
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<p class="MsoNormal"><span lang="EN-US">We transfected H9C2 cell with target plasmids, BBa_K2865006 and BBa_K2865005, via liposome transfection. After 48 hours’ expression, we detected the cells using fluorescence microscope. And we have taken clearly photos. We all know that the CMV promoter is widely and commonly used in plasmids or viruses designed for gene expression in mammalian cells since it is a very strong promoter and drives constitutive expression of genes under its control. And we have to admit that the transfection efficiency is low. </span></p>
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<p class="MsoNormal" style="text-indent: 0cm;"><span lang="EN-US">&nbsp;</span></p>
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[[File:T--SMMU-China--cep.png|500px|thumb|center|Figure 2 Expression of  CMV-eGFP-poly(A) was taken under fluorescent microscope and light microscope.]]
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[[File:T--SMMU-China--CMV without.png|300px|thumb|center|Fig.3 Expression of CMV-EGFP 48 h later without adding]]
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[[File:T--SMMU-China--CMV et-1+Ang 2.png|550px|thumb|center|Figure.4 Expression of CMV-EGFP 48 h after the administration of AngⅡand ET-1 under fluorescent microscope. (A) was cells treated with 10<sup>-7</sup> mol/L AngⅡ and (B) was cells treated with 10<sup>-7</sup> mol/L ET-1.]]
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<p class="MsoNormal" style="text-indent: 0cm;"><span lang="EN-US">&nbsp;</span></p> CMV promoter-eGFP-poly(A) served as the positive control. It contains the CMV promoter and eGFP reporter.
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 12:01, 15 October 2018


CMV promoter-eGFP-poly(A)

This part contains CMV promoter, eGFP reporter in RFC25 format(BBa_K2865005), and SV40 polyA terminator (BBa_K2865004). This part is constructed as a control with our part BNP promoter-eGFP-poly(A) (BBa_K2865008).

 

Fig.1 the construction of part

Experiment

Fluorescence microscope

We transfected H9C2 cell with target plasmids, BBa_K2865006 and BBa_K2865005, via liposome transfection. After 48 hours’ expression, we detected the cells using fluorescence microscope. And we have taken clearly photos. We all know that the CMV promoter is widely and commonly used in plasmids or viruses designed for gene expression in mammalian cells since it is a very strong promoter and drives constitutive expression of genes under its control. And we have to admit that the transfection efficiency is low.

 

Figure 2 Expression of CMV-eGFP-poly(A) was taken under fluorescent microscope and light microscope.
Fig.3 Expression of CMV-EGFP 48 h later without adding
Figure.4 Expression of CMV-EGFP 48 h after the administration of AngⅡand ET-1 under fluorescent microscope. (A) was cells treated with 10-7 mol/L AngⅡ and (B) was cells treated with 10-7 mol/L ET-1.

 

CMV promoter-eGFP-poly(A) served as the positive control. It contains the CMV promoter and eGFP reporter.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1430
  • 1000
    COMPATIBLE WITH RFC[1000]