Difference between revisions of "Part:BBa K4046610"

Latest revision as of 01:04, 22 October 2021

hUBC-Kozak-tdTomato

tdTomato is an important red fluorescence protein that has numerous applications in cell imaging and bio-sensing. When produced by the cell, it can generate red light upon excitation with yellow wavelength light. By placing it within a reporter construct, tdTomato and its associated fluorescence can be used to dynamically measure the levels of the compound of interest within the cell. This reporter can be used as a fluorescence-based constitutive reporter to quantitatively and non-invasively assess cell behavior.

To test the gene expression of hUBC-tdTomato, we transfected a plasmid containing this construct into HEK293T cells, and cells were imaged using fluorescence microscopy two days later. Overall, the results demonstrate that the tdTomato gene behaves as expected, as it shows fluorescence behavior when excited.

Figure 1: Fluorescence microscopy imaging of HEK293T cells transfected with hUBC-tdTomato plasmid

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal PstI site found at 759
Illegal PstI site found at 1485
12
INCOMPATIBLE WITH RFC[12]
Illegal PstI site found at 759
Illegal PstI site found at 1485
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal PstI site found at 759
Illegal PstI site found at 1485
25
INCOMPATIBLE WITH RFC[25]
Illegal PstI site found at 759
Illegal PstI site found at 1485
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K4046610 short</partinfo>
-This is a constitutive reporter consisting of the hUBC promoter followed by the gene coding for the tdTomato red fluorescence protein. In between these two sequences is the Kozak sequence, which provides a start point for eukaryotic translation. This construct was added to a pcDNA5 plasmid and was created as one of our constitutive reporters; it will be used as a construct into which we insert our <i>dhdO</i> binding sequences in order to build a dynamic reporter for the compound D-2-HG.
+tdTomato is an important red fluorescence protein that has numerous applications in cell imaging and bio-sensing. When produced by the cell, it can generate red light upon excitation with yellow wavelength light. By placing it within a reporter construct, tdTomato and its associated fluorescence can be used to dynamically measure the levels of the compound of interest within the cell. This reporter can be used as a fluorescence-based constitutive reporter to quantitatively and non-invasively assess cell behavior.
+To test the gene expression of hUBC-tdTomato, we transfected a plasmid containing this construct into HEK293T cells, and cells were imaged using fluorescence microscopy two days later. Overall, the results demonstrate that the tdTomato gene behaves as expected, as it shows fluorescence behavior when excited.
+[[File:T--Duke--tdTomato.png|800 px]] <br>
+<b><font size="+1">Figure 1: Fluorescence microscopy imaging of HEK293T cells transfected with hUBC-tdTomato plasmid</font></b>
 <!-- Add more about the biology of this part here