Jia Xu et al. Cell 2013 14 J23100 J23101 J23105 J23110 12 10 - - PowerPoint PPT Presentation

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Jia Xu et al. Cell 2013 14 J23100 J23101 J23105 J23110 12 10 - - PowerPoint PPT Presentation

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Jia Xu et al. Cell 2013 14 J23100 J23101 J23105 J23110 12 10 RFU (A.U.) 8 6 4 2 0 -7 -6 -5 -4 -3 log10 Kana (umol/L) Mut-3 Mut-8 1.42 1.34 1.40 1.32 1.38 1.30 RFU (A.U.) RFU (A.U.) 1.36 1.28 1.34 1.26 1.32 1.24

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Jia Xu et al. Cell 2013

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2 4 6 8 10 12 14 log10 Kana (umol/L) RFU (A.U.) J23100 J23101 J23105 J23110

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  • 6.0
  • 5.5
  • 5.0
  • 4.5
  • 4.0
  • 3.5

1.24 1.26 1.28 1.30 1.32 1.34

Mut-3

log10 [Kana] (umol/L) RFU (A.U.)

  • 6.0
  • 5.5
  • 5.0
  • 4.5
  • 4.0
  • 3.5

1.32 1.34 1.36 1.38 1.40 1.42

Mut-8

log10 [Kana] (umol/L) RFU (A.U.)

  • 6.0
  • 5.5
  • 5.0
  • 4.5
  • 4.0
  • 3.5

1.30 1.32 1.34 1.36 1.38 1.40

Mut-10

log10 [Kana] (umol/L) RFU (A.U.)

  • 6.0
  • 5.5
  • 5.0
  • 4.5
  • 4.0
  • 3.5

1.28 1.30 1.32 1.34 1.36 1.38 1.40

Mut-11

log10 [Kana] (umol/L) RFU (A.U.)

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–

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  • 6.0
  • 5.5
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  • 4.5
  • 4.0

2 4 6 8 10 12 log10 Kana (umol/L) Expression level (A.U.) J23100 J23101 J23104 J23110 R1051 R0010

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2 4 6 8 10 12 14 log10 Kana (umol/L) RFU (A.U.) J23100 J23101 J23105 J23110

ALeader (w.t.) ALeaderT

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  • 6.0
  • 5.5
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  • 4.5
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pSB1C3-J23100-loci-X-mRFP1

log10 Kana (umol/L) Expression level Aleader AleaderT AleaderT-BCD

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  • 6.0
  • 5.5
  • 5.0
  • 4.5
  • 4.0
  • 3.5

5 10 15 log10[Kana] (mol/L) RFU (A.U.) Aleader AleaderT

  • 6.0
  • 5.5
  • 5.0
  • 4.5
  • 4.0
  • 3.5

5 10 15 log10[Kana] (mol/L) RFU (A.U.) Aleader AleaderT

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Calculated by RNA fold SPR signal Data Model Fits Experiment Conformati

  • n

∆G˚ B (Binding)

  • 7.8

A (Free)

  • 4.3
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J S Paige et al. Science 2012

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 –  –

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Interacted CC JK

B0015 Terminator Efficiency

Terminator Efficiency 0.0 0.2 0.4 0.6 0.8 1.0 Interacted Normal

Test Input Test Output

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dA dt =kfA-(k1+k3+km)A+k1'B+k3'C dB dt =kfB+k1A-(k1'+k4'+k2L+km)B+k2'BL+k4C dBL dt =k2LB-(k2'+km)BL dC dt =kfC+k3A+k4'B-(k3'+k4+km)C dAm dt =kMAA-(k1+k3+kdMA)Am+k1'Bm+k3'Cm dBm dt =kMBB+k1Am-(k1'+k4'+k2L+kdMB)Bm+k2'BLm+k4Cm dBLm dt =kMBBL+k2LBm-(k2'+kdMB)BLm dCm dt =kMCC+k3Am+k4'Bm-(k3'+k4+kdMC)Cm dT dt =ktAA+ktB(B+BL)+ktCC-kdTT dP dt =kpAAm+kpB(Bm + BLm)+kpCCm-kdPP

C (Artificial Free)

  • 11.80

Conformation ∆G˚ B (Natural Binding)

  • 7.8

A (Natural Free)

  • 4.3

Partitioning Parameter No ribosome Influence K1 1.5234 K3 0.4309 K4 0.6564