Part:BBa_K1316016
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 <a href="http://2014.igem.org/Team:TU_Delft-Leiden/Project/Life_science/curli/characterisation" style="text-decoration: none"" target="_blank">conductive curli characterisation</a> on our wiki page!
Figure 1. Cells carrying the BBa_K1316016 construct observed under the confocal microscope scanning for Green fluorescence
Figure 2. Empty cells (not carrying the BBa_K1316016 construct) observed under the confocal microscope scanning for Green fluorescence. No fluorescent cells were observed
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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