Part:BBa_K4628008

Pikm-1*(Enhancer)

Transforming the ID sequence of the original Pikm-1 to an Enhancer nanoantibody targeting GFP, thereby artificially altering its immune inducible stimuli.

Overview of Pi-km

Pi-km is a major resistance allele on the rice blast resistance site : Pik, consisting of two closely linked NB-LRR genes with independent functions (Pikm1-TS and Pikm2-TS). Pi-km mediated rice blast resistance requires the joint action of Pikm1-TS and Pikm2-TS. When only one of Pikm1-TS or Pikm2-TS is present, rice exhibits susceptibility.

Introduction to Pikm-1

Pikm-1 is the component responsible for serving as the primary immune receptor, consisting of the N-terminal CC domain, the central NB-ARC domain, and the C-terminal LRR domain. Generally, we know that the N-terminal domain is involved in immune recognition and activation, while the central domain is an switch controlling the activity of the entire NLR protein through ADP/ATP exchange. At last, the C-terminal domain achieves fine regulation of protein activity through flexible folding.

Introduction to ID sequence

Meanwhile, it is found that Pikm-1 molecule has a special protein domain called the ID sequence. This sequence can directly bind to the effector secreted by rice blast fungus, thereby achieving specific immunity against rice blast. And the existence of ID sequence also provides a theoretical basis for our engineering modification of NLR molecules to change their resistance spectrum.

Introduction to Pikm-1*(Enhancer)

Pikm-1*(Enhancer) is such a molecular whose ID sequence has been replaced with a nanoantibody called Enhancer targeting GFP. This enables the modified NLR molecule to generate an immune response upon GFP stimulating, demonstrating a possibility of artificially modifying the NLR recognition spectrum.

Figure.1 Schematic diagram of modifying Pikm-1 molecule^[1]

Reference

[1] Kourelis J, Marchal C, Posbeyikian A, Harant A, Kamoun S. NLR immune receptor-nanobody fusions confer plant disease resistance. Science. 2023 Mar 3;379(6635):934-939.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 1496
  Illegal SpeI site found at 2070
  Illegal SpeI site found at 2076
  Illegal PstI site found at 2289
  Illegal PstI site found at 2505
  Illegal PstI site found at 2520
  Illegal PstI site found at 2589
  Illegal PstI site found at 3033
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 3619
  Illegal SpeI site found at 2070
  Illegal SpeI site found at 2076
  Illegal PstI site found at 2289
  Illegal PstI site found at 2505
  Illegal PstI site found at 2520
  Illegal PstI site found at 2589
  Illegal PstI site found at 3033
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2725
  Illegal BamHI site found at 2350
  Illegal XhoI site found at 3183
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 1496
  Illegal SpeI site found at 2070
  Illegal SpeI site found at 2076
  Illegal PstI site found at 2289
  Illegal PstI site found at 2505
  Illegal PstI site found at 2520
  Illegal PstI site found at 2589
  Illegal PstI site found at 3033
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 1496
  Illegal SpeI site found at 2070
  Illegal SpeI site found at 2076
  Illegal PstI site found at 2289
  Illegal PstI site found at 2505
  Illegal PstI site found at 2520
  Illegal PstI site found at 2589
  Illegal PstI site found at 3033
  Illegal NgoMIV site found at 721
  Illegal NgoMIV site found at 736
  Illegal NgoMIV site found at 2104
  Illegal NgoMIV site found at 3201
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 2334
  Illegal SapI site found at 1272
  Illegal SapI.rc site found at 531
  Illegal SapI.rc site found at 3242