Part:BBa_K208045
Lac Promoter/RBS/GFP/Term
This is a composite part using the GFP Silver-fusion compatible BioBrick K208000. This part is functional and cells harboring this part glow very brightly green when exposed to UV light. It functions as a reporter gene. This particular part is an example of how Silver-fusion compatible parts can still be used for non-fusion applications. Please see the original pages for the parts used in this composite for more details on their functionality.
Usage and Biology
This part can be used as a reporter gene.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 230
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 749
Illegal XhoI site found at 650 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
GFP activity is not observed in cells without the BBa K208045 BioBrick. GFP activity is also not seen in BBa K208045 containing XL1-blue cells (which have a functional lac repressor) and no IPTG. E. coli cells, like TOP10, do not have a functional lac repressor so GFP is expressed during growth. These results demonstrate the functionality of this BBa K208045 composite BioBrick.
Sodium dodecyl sulfate polyacrylamide gel electrophoresis was used to analyze the protein content in transformed E. coli. As a positive control, E. coli containing the Lac/RBS/GFP/Terminator (BBa_K208045) construct were sonicated and centrifuged (see Figure 6). Additionally, E. coli cells containing an individual BioBrick part (BBa_B0015) were analyzed as a negative control. The resulting gel was stained with coomassie blue and is shown. The bright band at 27 kD in the GFP+ sample corresponds to the GFP protein. The absence of this band in the GFP- sample further reinforces the functionality of the GFP construct.
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