Difference between revisions of "Part:BBa K1119008"
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In characterization of Mitochondrial Leader Sequence ([[Part:BBa_K1119001|BBa_K1119001]]), negative control was made by GFP generator that does not contains the CDS of MLS ([[Part:BBa_K1119008|BBa_K1119008]]). The translation unit was driven by CMV promoter ([[Part:BBa_K1119006|BBa_K1119006]]) and terminated by hGH polyA signal ([[Part:BBa_K404108|BBa_K404108]]).  
 
In characterization of Mitochondrial Leader Sequence ([[Part:BBa_K1119001|BBa_K1119001]]), negative control was made by GFP generator that does not contains the CDS of MLS ([[Part:BBa_K1119008|BBa_K1119008]]). The translation unit was driven by CMV promoter ([[Part:BBa_K1119006|BBa_K1119006]]) and terminated by hGH polyA signal ([[Part:BBa_K404108|BBa_K404108]]).  
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===Result===
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In the characterization of CMV promoter ([[Part:BBa_K1119006|BBa_K1119006]]) , the coding sequence of CMV promoter was assembled with GFP reporter ([[Part:BBa_K648013|BBa_K648013]]) and hGH polyA terminator ([[Part:BBa_K404108|BBa_K404108]]) using Freiburg’s RFC25 format.
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The pCMV-GFP was then transfected into HEK293FT cells and in vivo green fluorescence signal was observed under confocal microscope.
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The positive control was pEGFP-N1 (Clontech) that contains CMV promoter and EGFP reporter. A negative control was made by GFP generator that does not contain the CMV promoter.
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The [http://2013.igem.org/Team:Hong_Kong_HKUST/characterization detailed protocol] of our characterization can be found in HKUST iGEM 2013 Wiki.
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[[File:Final CMV annotated no ABC.jpg|600px|thumb|center|'''Figure 1. CMV promoter drives expression of GFP.''' HEK cells transfected with pCMV-GFP expressed GFP Signal. HEK cells transfected with the commercial pEGFP-N1 showed similar results, while the negative control GFP without any promoter was not expressed.  ]]
  
  

Revision as of 11:53, 26 September 2013

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In characterization of Mitochondrial Leader Sequence (BBa_K1119001), negative control was made by GFP generator that does not contains the CDS of MLS (BBa_K1119008). The translation unit was driven by CMV promoter (BBa_K1119006) and terminated by hGH polyA signal (BBa_K404108).

Result

In the characterization of CMV promoter (BBa_K1119006) , the coding sequence of CMV promoter was assembled with GFP reporter (BBa_K648013) and hGH polyA terminator (BBa_K404108) using Freiburg’s RFC25 format.

The pCMV-GFP was then transfected into HEK293FT cells and in vivo green fluorescence signal was observed under confocal microscope.

The positive control was pEGFP-N1 (Clontech) that contains CMV promoter and EGFP reporter. A negative control was made by GFP generator that does not contain the CMV promoter.

The [http://2013.igem.org/Team:Hong_Kong_HKUST/characterization detailed protocol] of our characterization can be found in HKUST iGEM 2013 Wiki.

Figure 1. CMV promoter drives expression of GFP. HEK cells transfected with pCMV-GFP expressed GFP Signal. HEK cells transfected with the commercial pEGFP-N1 showed similar results, while the negative control GFP without any promoter was not expressed.


Sequence and Features Status: 500 Content-type: text/html

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