Difference between revisions of "Part:BBa K1316016"

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For more information about the characterisation of this construct visit the  
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For more information about the characterisation of this construct visit the conductive curli characterisation on our wiki page! (http://2014.igem.org/Team:TU_Delft-Leiden/Project/Life_science/curli/characterisation)
<a href="http://2014.igem.org/Team:TU_Delft-Leiden/Project/Life_science/curli/characterisation" style="text-decoration: none"" target="_blank"><font color="#0080FF" size="3">conductive curli characterisation</font></a>
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on our wiki page!
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Revision as of 11:19, 13 October 2014

constitutively expressed eGFP under the control of J23110 Anderson promoter

Constitutively expressed Enhanced Green Fluorescen Protein (eGFP). eGFP is more intense than normal GFP. The purpose of this construct is to be able to easily detect cells attached to a surface, therefore detecting biofilm formation cells. For our project that would be used for detecting curli-forming cells.

Characterisation

eGFP expression was observed on the confocal microscope. All the cells carrying the BBa_K1316016 construct showed green fluorescence corresponding to eGFP (Excitation wavelength: 488nm; Emission wavelength: 508nm), whereas empty cells not carrying the BBa_K1316016 construct showed no fluorescence.

For more information about the characterisation of this construct visit the conductive curli characterisation on our wiki page! (http://2014.igem.org/Team:TU_Delft-Leiden/Project/Life_science/curli/characterisation)


Delft2014_Gfp_fluorescent.jpg

Figure 1. Cells carrying the BBa_K1316016 construct observed under the confocal microscope scanning for Green fluorescence

TUDelft_DcsgB_induced.jpg

Figure 2. Empty cells (not carrying the BBa_K1316016 construct) observed under the confocal microscope scanning for Green fluorescence. No fluorescent cells were observed

TUDelft_DcsgB_induced_Brightfield.jpg

Figure 3. Same field of Figure 2 but scanned with the Bright-Field mode to prove that, even though there was no fluorescence observed there were indeed cells present. Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]