Part:BBa_K1624002
LacO
Lac operator sequence, involved in transcriptional regulation of the lac operon. Can be placed between promoter and RBS sequence to regulate expression of gene by induction.
Sequence from lacO in BL21(DE3) strain.
Characterization in vivo
The lacO sequence was placed between a T7 promoter and sfGFP gene (BBa_K1624004), regulating GFP expression. The lacO sequence was also in the E. coli genome regulating expression of T7 RNA polymerase. The experimental construct was placed in a pCDFDuet-1 vector and transformed into BL21(DE3) cells. Cells were grown overnight. A 1:100 dilution was made, and cultures were grown with 20% glucose to 0.6 OD. At 0.6 OD, cells were resuspended in IPTG and induced for 2 hours. After induction, cells were washed and resuspended in PBS buffer. Fluorescence was then measured at 600 nm for 25 flashes. Bars show average of triplicates. Error bars show standard deviation.
The graph above shows a significantly higher expression of GFP with IPTG than without. The lacO sequence successfully regulating expression of T7 RNA polymerase in the genome and sfGFP in the plasmid.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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