BioSketch Components

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{For each component, identify and describe the gene, the promoter regulated by the gene, the induction time course, and the relevant references.}

A

• UV-response proteins
  • RecA
    • activated by ssDNA (Kobayashi et al., 2004; Yasuda, 1998)
    • cleaves cI repressors (Kobayashi et al., 2005; Yasuda, 1998)
  • cI
    • cleaved by active RecA (Kobayashi et al., 2004; Yasuda, 1998)
    • represses:
      • P_L* promoter (promotes lacI; Kobayashi et al., 2004)
      • λP_R promoter (Elowitz & Leibler, 2000)
      • P^λ- promoter (Guet et al., 2002)
    • activates:
      • P^λ+ promoter (Guet et al., 2002)
  • 434 repressor
    • cI analog from lambdoid phage 434
    • represses P₄₃₄ promoter
• DNA Damage response promoter?
• Promoter: blocked by B

B

• Temperature Sensitive Repressors
  • cI (Gardner et al, 2000)
  • lacI(ts) (Hasan & Szybalski, 1995; Yabuta et al., 1995; Chao et al., 2002; Andrews et al., 1996; Bukrinsky et al., 1988)
    • represses:
      • lacO operator (Hasan & Szbalski, 1995; Wang et al., 2004)
      • P_LlacO1 promoter (Elowitz & Leibler, 2000: reported for WT LacI)
      • P_trc promoter (promotes cI; Kobaysahi et al., 2005: reported for WT LacI)
      • P^L1 promoter (Guet et al., 2002: reported for WT LacI)
      • P^L2 promoter (Guet et al., 2002: reported for WT LacI)
    • strain needs to be deficient in endogenous lacI:
      • LBG1081 (Hasan & Szbalski, 1995)
      • MYW3110 (Yabuta et al., 1995)
  • mu c (Vogel et al., 1991)
  • tet reressor (Wissmann, 1991)
  • araC (Irr & Englesberg, 1971)
• Heat-shock response promoter?
• Promoter: blocked by A

Reporter

• mCherry - fast maturation fluorophore, appears purple in visible light
• Bacterial alpha-beta luciferase fusion protein - cannot fold at high temperatures
• Promoter: blocked by A