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Lac Operon & Mutants

Lac Operon & Mutants. lac Operon. lacI. CAP. p. o. lacZ. lacY. lacA. Acetylase. Lac Repressor 38,00 kD. allolactose. Lac Permease , ( symporter ) 1.5 kb mRNA, 46,500 kD. β - galactosidase , 3.1 kb mRNA, 120 kD glc -gal  glc + gal β-1-4 lactose  β-1-4 allolactose.

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Lac Operon & Mutants

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  1. Lac Operon & Mutants

  2. lacOperon lacI CAP p o lacZ lacY lacA Acetylase Lac Repressor 38,00 kD allolactose Lac Permease, (symporter) 1.5 kb mRNA, 46,500 kD β-galactosidase, 3.1 kb mRNA, 120 kD glc-gal  glc + gal β-1-4 lactose  β-1-4 allolactose

  3. Wild type Media with X-gal + IPTG Media with X-gal - IPTG *Uptake of X-gal and IPTG controlled by Lac Permease *Small amount of Lac Permease present in membrane at all times *IPTG not degraded

  4. Wild type Media with ONPG, toluene, + IPTG Media with ONPG, toluene, - IPTG *X-gal and IPTG move freely into cells because of toluene

  5. What would the mutants be like?

  6. lacZ Mutants Media with X-gal + IPTG Media with X-gal - IPTG Media with ONPG, toluene, + IPTG Media with ONPG, toluene, - IPTG

  7. What would the mutants be like?

  8. lacY Mutants Media with X-gal + IPTG Media with X-gal - IPTG Media with ONPG, toluene, + IPTG Media with ONPG, toluene, - IPTG

  9. What would the mutants be like?

  10. lacO Mutants Media with X-gal + IPTG Media with X-gal - IPTG Media with ONPG, toluene, + IPTG Media with ONPG, toluene, - IPTG

  11. What would the mutants be like?

  12. What would the mutants be like?

  13. lacIc Mutants and lacIS Mutants Media with X-gal + IPTG Media with X-gal - IPTG Media with ONPG, toluene, - IPTG Media with ONPG, toluene, + IPTG

  14. Table 4 – LacI is a trans acting factor

  15. What about cAMP and CRP? cya AMP ------- AdenylateCyclase --------- CRP – cAMP Receptor Protein Or CAP – Catabolite Activator Protein crp

  16. Let’s take a closer look

  17. What would the mutants be like?

  18. E. coli on minimal medium with Glycerol / IPTG

  19. What effect would cAMP have?

  20. E. coli on minimal medium with Glycerol / IPTG

  21. What would the mutants be like?

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