Functional reconstitution of a bacterial CO2 concentrating mechanism in E. coli.
Flamholz AI, Dugan E, Blikstad C, Gleizer S, Ben-Nissan R, Amram S, Antonovsky N, Ravishankar S, Noor E, Bar-Even A, Milo R, Savage D
Elife. 2020 Oct 21;9. pii: 59882. doi: 10.7554/eLife.59882.
(Link opens in a new window)
PubMed
(Link opens in a new window)
Article
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 162688 | pFE-sfGFP | pFE vector control expressing sfGFP |
| 162689 | pFA-sfGFP | pFA vector control expressing sfGFP |
| 162690 | pFA-DAB2 | Expresses DabAB2 inorganic carbon transport complex |
| 162691 | pFA-DAB1 | Expresses DabAB1 inorganic carbon transport complex |
| 162692 | p1A | Expresses H. neapolitanus CbbLS and S. elongatus Prk. Rescues CCMB1 in M9 glycerol at 10% CO2 |
| 162693 | p1A CbbL- | p1A negative control expressing inactive CbbLS (carbamylated K194M mutation) and Prk |
| 162694 | p1A Prk- | p1A negative control expressing CbbLS and inactive Prk (Prk K20M S21A) |
| 162695 | pFE-Rrub-FII-prk | Expresses R. rubrum Form II rubisco and S. elongatus Prk |
| 162696 | pFE-Reu-FI-prk | Expresses R. eutropha Form IC rubisco and S. elongatus Prk |
| 162697 | pFE-Rs-FI-prk | Expresses R. spaeroides Form IC rubisco and S. elongatus Prk |
| 162698 | pFE-Selo-FI-prk | Expresses S. elongatus Form IB rubisco and S. elongatus Prk |
| 162699 | pCB | Expresses pHnCB10 derived carboxysome operon and prk |
| 162703 | pCB’ | pCB reconstructed after selection for CCMB1 growth on glycerol under ambient air |
| 162705 | pCB’ CsoS2 ΔNTD | Carboxysome operon with truncated CsoS2 (deletion of amino acids 2-256), prk; Deletion of rubisco-binding N-terminus of CsoS2 |
| 162707 | pCB’ CbbL Y72R | Carboxysome operon with point mutation to CbbL, prk; Substitution of CsoS2-binding tyrosine with arginine prevents rubisco encapsulation creating rubisco-less carboxysomes |
| 162708 | pCCM | Second CCM operon cloned from H. neapolitanus; pFA expressing the remaining 11 genes in the H. neapolitanus CCM cluster |
| 162709 | pCCM’ | pCCM reconstructed after selection for CCMB1 growth on glycerol under ambient air |
| 162710 | pCCM’ ΔacRAF | Second CCM operon lacking acRAF; Deletion of putative rubisco chaperone, acRAF |
| 162711 | pCCM’ CbbQ- | Second CCM operon with inactive CbbQ (K46A, E107Q) rubisco activase |