Part:BBa_K2958002
Lac Promoter + RFP
Red fluorescent protein (BBa_E1010) regulated by the lac promoter (BBa_R0010) with medium rbs BBa_B0034.
Description:
Red Fluorescent Protein (RFP) is a reporter gene used to confirm the expression of a circuit. RFP is easily visible, as the cells will glow red if they are properly expressed. This circuit is regulated by the Lac Promoter. If the Lac repressor is not used with the lac promoter, then the promoter will act as a constitutive promoter and the red fluorescence will always be expressed. If this circuit is used with the Lac Repressor, RFP will only be expressed when lactose or IPTG are present.
Characterization:
Figure 1.Gel electrophoresis ran at 110V for 30 minutes. Lane 1 is APEX 500 bp ladder, lane 2 is PCR purified LacP+RFP at the correct basepair length (924 bp), lane 3 is PCR purified proinsulin at the correct basepair length (1068 bp), NL 5.50 Single Chain Proinsulin at the correct basepair length (not used for this project; 1002 bp), and lane 4 is long lasting single chain insulin at the correct basepair length (1002 bp). This gel electrophoresis result supports that the LacP+RFP gene block was properly PCR'd. Therefore, the RFP expression observed in Figure 2 and Figure 3 can be attributed to the expression of this DNA sequence.
Figure 2. Transformation of Proinsulin + RFP Construct (BBa_K2958013)into BL21 cells glowing red due to successful RFP expression.
Figure 3. Overnight of extracted protein from Proinsulin + RFP Construct (BBa_K2958013) that remained red due to the LacP + RFP.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 781
Illegal AgeI site found at 893 - 1000COMPATIBLE WITH RFC[1000]
| None |