Part:BBa_K3900009
NOS Plant Promoter
NOS Plant Promoter
Thessaly’s 2022 Contribution
Overview
The Nopaline Synthase Promoter (or NOS Promoter) is a regulatory element of bacterial origin that can drive expression in plant tissues. Normally, it is used for generation of an unusual compound, called Nopaline, that, when secreted into the environment, can be used by Agrobacterium tumefaciens as a nutrient source [1]. In synthetic biology, wherein transgene expression can be achieved through the creation of novel devices, pNOS has been established as a Phytobrick for the ability to constitutively express genes in Plant Tissues [2]. In comparison to the obvious counterpart, the Cauliflower Mosaic Virus 35s Promoter (CaMV 35s Promoter), it leads to lower levels of expression [3], desirable in conditions where increased transgene presence results in toxicity to the host or a non-responsive device. iGEM Thessaly 2022 has managed to confirm the functionality of this part by designing a Reporter System composed of the basic parts pNOS:mVenus:tNOS (BBa_K4213029).
Experimental Design and Results
We selected Ν. benthamiana since it allows for high and rapid expression of transgenes, by agroinfiltration, as the species is quite susceptible to plant viral vectors [4]. N. benthamiana leaf samples are also able to be observed under UV lumination for expression of fluorescent proteins [4]. As for the cloning method, we opted for the Golden Braid for its simplicity, effectiveness and rapidness [5] and managed to confirm assembly with the help of diagnostic digestion.
Following that, came agroinfiltration and observation under the confocal microscope 4 days post infiltration (dpi) [6], whereupon we confirmed the functionality of the Transcriptional Unit (TU).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 114
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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