In our project, our team had the goal of genetically engineering Chlamydomonas reinhardtii to increase its ability to sequester phosphorous, and to increase its uptake of nitrate by converting nitrous oxide to dinitrogen gas.
To accomplish this, our team developed 3 new simple parts. We utilized additional simple parts from the Chlamdydomonas reinhardtii MoClo toolkit (Crozet et al. 2016), which have been documented by other iGEM Teams, along with a simple part for the Psr1 gene from the Alison Baker Lab (Slocombe et al. 2023). This led to the generation of 8 new composite parts. A brief description of these parts is in the table below.
For more information, including a description of which simple parts were used to assemble each composite part, please visit the part’s Registry page!
part | Chlamy MoChlo Toolkit # | Registry Number | iGEM Team/Collaborator | Part Description/Purpose |
---|---|---|---|---|
nanoLuc | PCM0-104 | K2619004 | iGEM18_XJTLU-CHINA | luciferase reporter |
aadA CDS | PCM0-076 | K3002000 | iGEM19_TU_Kaiserslautern | spectinomycin resistance |
Psad Promoter+5' UTR | PCM0-016 | K3002001 | iGEM19_TU_Kaiserslautern | Strong promoter in C. reinhardtii |
Psad Terminator | PCM0-114 | K3002002 | iGEM19_TU_Kaiserslautern | terminator in C. reinhardtii |
Btub promoter+5' UTR | PCM0-021 | K3002011 | iGEM19_TU_Kaiserslautern | Strong promoter in C. reinhardtii |
Btub Terminator | PCM0-118 | K3002012 | iGEM19_TU_Kaiserslautern | terminator in C. reinhardtii |
mCherry | PCM0-108 | K4770013 | iGEM23_Barcelona-UB | fluorescent reporter |
NosZ P. stutzeri | K5078000 | new | nitrous oxide reductase from Pseudomonas stutzerii | |
NosZ D. denitirficans | K5078001 | new | nitrous oxide reductase from Dechloromonas denitrificans | |
Ptc amiRNA | K5078002 | new | encodes miRNA against Ptc mRNA | |
Psr gDNA | K5078008 | provided by Alison Baker Lab (Slocombe et al. 2023) | Phosphate starvation response 1 transcription factor | |
pL1-aadA | K5078004 | new | Composite part - provide C. reinhardtii with resistance to spectinomycin | |
pL1 NosZ (Pstu)-mCherry | K5078005 | new | Composite part - allow C. reinhardtii to convert nitrous oxide to dinitrogen gas, tagged with mCherry reporter | |
pL1 NosZ (D denit)-mCherry | K5078006 | new | Composite part - allow C. reinhardtii to convert nitrous oxide to dinitrogen gas, tagged with mCherry reporter | |
pL1 ptc amiRNA | K5078007 | new | Composite part - reduce expression of Ptc mRNA | |
pL1 Psr1-nanoLuc | K5078003 | new | Composite part - upregulate phosphate starvation response in C. reinhardtii, tagged with nanoLuc reporter | |
pL2 -aadA-nosZ(P.stu)-Ptc amiRNA-Psr | K5078009 | new | Composite part - combination of above pL1 functions | |
pL2 -aadA-nosZ(D.denit)-Ptc amiRNA-Psr | K5078010 | new | Composite part - combination of above pL1 functions | |
pL2-aadA-Psr | K5078011 | new | Composite part - combination of above pL1 functions |