scholarly journals A Low-CO2-Inducible Gene Encoding an Alanine:[alpha]-Ketoglutarate Aminotransferase in Chlamydomonas reinhardtii

1996 ◽  
Vol 112 (2) ◽  
pp. 677-684 ◽  
Author(s):  
Z. Y. Chen ◽  
M. D. Burow ◽  
C. B. Mason ◽  
J. V. Moroney
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Gene Encoding ◽  
Low Co2 ◽  
Inducible Gene

scholarly journals Rubisco Activase Is Required for Optimal Photosynthesis in the Green Alga Chlamydomonas reinhardtii in a Low-CO2 Atmosphere

2003 ◽  
Vol 133 (4) ◽  
pp. 1854-1861 ◽  
Author(s):  
Steve V. Pollock ◽  
Sergio L. Colombo ◽  
Davey L. Prout ◽  
Ashley C. Godfrey ◽  
James V. Moroney
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Green Alga ◽  
Rubisco Activase ◽  
Low Co2 ◽  
Co2 Atmosphere

Structure and expression of the gene encoding the periplasmic arylsulfatase of Chlamydomonas reinhardtii

1989 ◽  
Vol 218 (2) ◽  
pp. 229-239 ◽  
Author(s):  
Eugenio L. de Hostos ◽  
James Schilling ◽  
Arthur R. Grossman
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Gene Encoding

Properties of Selenium-Induced Glutathione Peroxidase in Low-CO2-Grown Chlamydomonas reinhardtii

1990 ◽  
pp. 3409-3412 ◽  
Author(s):  
Shigeru Shigeoka ◽  
Toru Takeda ◽  
Tsuyoshi Hanaoka ◽  
Akiho Yokota ◽  
Shozaburo Kitaoka ◽  
...  
Keyword(s):  
Glutathione Peroxidase ◽  
Chlamydomonas Reinhardtii ◽  
Low Co2

scholarly journals Successful Transient Expression of Cas9 and Single Guide RNA Genes in Chlamydomonas reinhardtii

2014 ◽  
Vol 13 (11) ◽  
pp. 1465-1469 ◽  
Author(s):  
Wenzhi Jiang ◽  
Andrew J. Brueggeman ◽  
Kempton M. Horken ◽  
Thomas M. Plucinak ◽  
Donald P. Weeks
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Transient Expression ◽  
Gene Knockout ◽  
Nuclease Activity ◽  
Gene Replacement ◽  
Target Site ◽  
Gene Encoding ◽  
Content Type ◽  
Guide Rna ◽  
Eukaryotic Organisms

ABSTRACT The clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system has become a powerful and precise tool for targeted gene modification (e.g., gene knockout and gene replacement) in numerous eukaryotic organisms. Initial attempts to apply this technology to a model, the single-cell alga, Chlamydomonas reinhardtii , failed to yield cells containing edited genes. To determine if the Cas9 and single guide RNA (sgRNA) genes were functional in C. reinhardtii , we tested the ability of a codon-optimized Cas9 gene along with one of four different sgRNAs to cause targeted gene disruption during a 24-h period immediately following transformation. All three exogenously supplied gene targets as well as the endogenous FKB12 (rapamycin sensitivity) gene of C. reinhardtii displayed distinct Cas9/sgRNA-mediated target site modifications as determined by DNA sequencing of cloned PCR amplicons of the target site region. Success in transient expression of Cas9 and sgRNA genes contrasted with the recovery of only a single rapamycin-resistant colony bearing an appropriately modified FKB12 target site in 16 independent transformation experiments involving >10 9 cells. Failure to recover transformants with intact or expressed Cas9 genes following transformation with the Cas9 gene alone (or even with a gene encoding a Cas9 lacking nuclease activity) provided strong suggestive evidence for Cas9 toxicity when Cas9 is produced constitutively in C. reinhardtii . The present results provide compelling evidence that Cas9 and sgRNA genes function properly in C. reinhardtii to cause targeted gene modifications and point to the need for a focus on development of methods to properly stem Cas9 production and/or activity following gene editing.

Identification and characterisation of a novel inorganic carbon acquisition gene, CIA7, from an insertional mutant of Chlamydomonas reinhardtii

2008 ◽  
Vol 35 (5) ◽  
pp. 373 ◽  
Author(s):  
Ruby A. Ynalvez ◽  
James V. Moroney
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Metal Ion ◽  
Inorganic Carbon ◽  
Antibiotic Resistance Gene ◽  
Inverse Pcr ◽  
Chloroplast Targeting ◽  
Calculated Molecular Mass ◽  
Prediction Program ◽  
Metal Transporter ◽  
Low Co2

Chlamydomonas reinhardtii is a unicellular eukaryotic alga which possesses a CO2-concentrating mechanism (CCM) that enables it to grow at low CO2 concentrations. Previously, insertional mutants were generated to enable isolation of inorganic carbon transporters and other proteins that might be essential for a functional CCM. These mutants have an antibiotic resistance gene that encodes a protein that binds to Zeocin inhibiting Zeocin’s DNA strand cleavage activity. The DNA flanking the BleR insert of one of the high CO2 requiring strains, named cia7, was cloned with inverse-PCR and sequenced. Sequence analysis showed homology to conserved bacterial proteins of unknown function, but there were no ESTs in this region of the genome. However, the presence of a gene was established by PCR and RLM-RACE. CIA7 was found to have four exons and the BleR insert was in the fourth exon. CIA7 encodes a protein of 104 amino acids with a calculated molecular mass of 11.3 kDa. Based on the ChloroP prediction program, the protein is predicted to have a chloroplast targeting signal. Complementation analyses results showed possible partially rescued mutants, and RNAi showed several transformants with a sick on low CO2 phenotype with reduced expression of CIA7. These results suggest that CIA7 is a gene that facilitates growth in C. reinhardtii under low CO2 conditions. One possible role of CIA7 would be in the delivery or storage of a metal ion. It may play a potential role as either a domain of a metal transporter or as a metallochaperone.

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