Part:BBa_K2036003
SnRK2.2,SNF1-related protein kinase 2.2
Together with SnRK2.2, key component and activator of the abscisic acid (ABA) signaling pathway that regulates numerous ABA responses, such as seed germination, Pro accumulation, root growth inhibition, dormancy and seedling growth, and, to a lesser extent, stomatal closure.
We apply SnRK2.2 into our eukaryotic filter. In our system, when signal on is present, SnRK2.2 express and phosphorylate ABF2 to activate the promoter RD29A form positive feedback. At the same time, ABF2 triggers the stable production of target gene.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 3
Illegal BamHI site found at 660 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Protein expression
Pichia pastoris are typical Eukaryotic expression host, but expressing plant genes are still a big challenge.So we firstly tested whether our three interest genes can be efficient expressed. We measured the secretion expression of PP2CA, ABF2 and SnRK2.2. We ran a SDS-PAGE of the culture supernatants of induced cells to identify the three recombinant proteins(Fig2). Compared to vector trasnfected cells, three kinds of recombinant protein plasmids trasnfected cells all expressed proteins at ~80kD under induction. The shift of protein bands suggested potential glycosylation modification.
Bi-stable function:
We constructed expression plasmid and submitted this part BBa_K2036030 to the registry.But due to the limited time,its function characterization is still under testing.However,our modeling simulation showed promising switch functions.See to Eukaryote circuit modeling.
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