Part:BBa_K4948033
PJEx-D-RBS-vsfGFP-0_lva
One of the plaamids comprising our biosensor, MoNitro, continuously produces GFP under normal conditions under the control of the PJEx-D promoter. However, when exposed to high concentrations of nitrate, the expression of the Mf_lon protease and EilR repressor, which are regulated by the nitrate inducible promoter, is overwhelmed, resulting in the degradation of lva-tagged GFP, and the transcription of the PJEx-D promoter is inhibited by EilR, resulting in low GFP levels.
Usage and Biology
Vector Map
Description
To monitor nitrate in streams, we first constructed a biosensor using molecular biology and genetic engineering at the laboratory level. We wanted our biosensor to show green at low nitrate conditions and red at dangerous levels of concentration. For this purpose, we used a strong constitutive promoter (pJEx-D-RBS) to show green color under normal conditions (BBa_K4948033). However, in order to degrade the already made GFP when nitrate is present in high concentration, a lva tag that can be recognized by protease was fused to the gfp. And to control at the transcriptional level, EilR, a repressor that negatively regulates the transcriptional activity of pJEx-D, was added to the nitrate induced system. At the laboratory level, we found that our biosensor worked smoothly enough to be distinguished by the eye in 6 hours.
At the laboratory level, we found that our biosensor worked smoothly enough to be distinguished by the eye in 6 hours. Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 199
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 199
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 199
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 199
Illegal NgoMIV site found at 451
Illegal AgeI site found at 557
Illegal AgeI site found at 971 - 1000COMPATIBLE WITH RFC[1000]
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