From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created two substitutions (V149Q and I282H) together in the Rubisco large subunit. This mutant (also named G149-282) represents one of 15 “groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created six substitutions (G168P, L326I, M349L, M375L, A398S, and C399V) together in the Rubisco large subunit. This mutant (also named G168-399) represents one of 15 “groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The mutant enzyme has decreases in CO2/O2 specificity and carboxylation catalytic efficiency (Lim and Spreitzer, unpublished). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created a V221C substitution in the Rubisco large subunit. This mutant (also named G221) represents one of 15 “groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created five substitutions (R305K, D470E, T471A, I472M, and K474T) together in the Rubisco large subunit. This mutant (also named G305-474) represents one of 15 “groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The mutant enzyme has a decrease in carboxylation catalytic efficiency (Lim and Spreitzer, unpublished). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created two substitutions (V391T and T428V) together in the Rubisco large subunit. This mutant (also named G391-428) represents one of 15 “groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The mutant enzyme has a decrease in carboxylase specific activity (Lim and Spreitzer, unpublished). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created four substitutions (G442N, D443E, V444I, and S447E) together in the Rubisco large subunit. This mutant (also named G442-447) represents one of 15 “groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The mutant has small decreases in photosynthetic growth and the amount of Rubisco holoenzyme (Lim and Spreitzer, unpublished). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created six substitutions (V30E, V31T, R32K, D86H, I105L, and C369V) together in the Rubisco large subunit. This mutant (also named AG30-32) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created eight substitutions (M42V, C53A, D86H, R305K, D470E, T471A, I472M, and K474T) together in the Rubisco large subunit. This mutant (also named AG42-53) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created six substitutions (V30E, V31T, R32K, M42V, C53A, and D86H,) together in the Rubisco large subunit. This mutant (also named AG86) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The mutant enzyme has a decrease in CO2/O2 specificity (Lim and Spreitzer, unpublished). The mutant strain has a small decrease in photosynthetic growth at 35 °C. It has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created seven substitutions (V30E, V31T, R32K, I105L, V149Q, I282H, and C369V) together in the Rubisco large subunit. This mutant (also named AG105-369) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created seven substitutions (I105L, V149Q, C256F, K258R, I265V, I282H, and C369V) together in the Rubisco large subunit. This mutant (also named AG149-282) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The mutant strain has small decreases in Rubisco holoenzyme and photosynthetic growth when grown at 35 °C (Lim and Spreitzer, unpublished). It has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created ten substitutions (G168P, I265V, L326I, V341I, M349L, M375L, V391T, A398S, C399V, and T428V) together in the Rubisco large subunit. This mutant (also named AG168-399) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Phenotype: requires acetate at 35 °C, temperature-conditional

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created four substitutions (V221C, C256F, K258R, and I265V) together in the Rubisco large subunit. This mutant (also named AG221) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The mutant enzyme has a decrease in CO2/O2 specificity (Lim and Spreitzer, unpublished). The mutant strain can grow on minimal medium at 25 °C, but dies on minimal medium at 35 °C, and has a decreased amount of Rubisco holoenzyme when grown at 35 °C with acetate medium in darkness. This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Phenotype: requires acetate, sensitive to light, spectinomycin resistant

This Rubisco rbcL knock-out mutant was created by Boon Hoe Lim in Spreitzer’s group by using aadA to replace chloroplast rbcL of rbcL-Penta/rbcS1-ABSO mt+ (CC-4848) (Spreitzer et al. 2005). The mutant lacks Rubisco large subunits, but is capable of expressing the ABSO small subunit, which was previously engineered by replacing the Chlamydomonas betaA-betaB loop with the loop from the small subunit of spinach Rubisco (Karkehabadi et al. 2005; Spreitzer et al. 2005). The mutant was created to investigate large-subunit amino acids that differ between Chlamydomonas and land plants, and that interact with small-subunit amino acids (Du et al. 2003). This strain has been cloned to homoplasmicity, and maintained with acetate medium in darkness since its isolation.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Karkehabadi S, Peddi SR, Anwaruzzaman M, Taylor TC, Cederlund A, Genkov T, Andersson I, Spreitzer RJ (2005) Chimeric small subunits influence catalysis without causing global conformational changes in the crystal structure of ribulose-1,5-bisphosphate carboxylase/oxygenase. Biochemistry 44:9851-9861

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL, RBCS1
  • Chromosome:
  • chloroplast,2

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆/rbcS1-ABSO mt+ (Spreitzer et al. 2005), Boon Hoe Lim in Spreitzer’s group created a V235I substitution in the Rubisco large subunit. This mutant (also named AG235) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The mutant enzyme has a decrease in CO2/O2 specificity (Lim and Spreitzer, unpublished). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL, RBCS1
  • Chromosome:
  • chloroplast,2

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆/rbcS1-ABSO mt+ (Spreitzer et al. 2005), Boon Hoe Lim in Spreitzer’s group created six substitutions (V149Q, V221C, C256F, K258R, I265V, and I282H) together in the Rubisco large subunit. This mutant (also named AG256) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The mutant strain has a small decrease in Rubisco holoenzyme when grown at 35 °C (Lim and Spreitzer, unpublished). It has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL, RBCS1
  • Chromosome:
  • chloroplast,2

From Robert J. Spreitzer, University of Nebraska, November 2014

Phenotype: requires elevated CO2 for photosynthetic growth

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆/rbcS1-SSAT pf2 mt+ (CC-4927), Boon Hoe Lim in Spreitzer’s group created 12 substitutions (V149Q, G168P, V221C, C256F, K258R, I265V, I282H, L326I, M349L, M375L, A398S, and C399V) together in the Rubisco large subunit. This mutant (also named AG265/SSAT) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The 12 substitutions in the large subunit must be complemented by the presence of the Arabidopsis small subunit (SSAT) to produce a functional Rubisco holoenzyme (Lim and Spreitzer, unpublished). The mutant strain requires 5% CO2 in air for photosynthetic growth (Genkov et al. 2010), and has a decrease in Rubisco holoenzyme stability (Lim and Spreitzer, unpublished). It has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Genkov T, Meyer M, Griffiths H, Spreitzer RJ (2010) Functional hybrid Rubisco enzymes with plant small subunits and algal large subunits: Engineered rbcS cDNA for expression in Chlamydomonas. J Biol Chem 285:19833-19841

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL, RBCS1
  • Chromosome:
  • chloroplast,2

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created eight substitutions (M42V, C53A, R305K, V341I, D470E, T471A, I472M, and K474T) together in the Rubisco large subunit. This mutant (also named AG305-474) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created 12 substitutions (G168P, R305K, L326I, V341I, M349L, M375L, A398S, C399V, D470E, T471A, I472M, and K474T) together in the Rubisco large subunit. This mutant (also named AG341) represents one of 15 “associated groups” of amino acids that differ between Chlamydomonas and land plants (Du et al. 2003). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Spreitzer et al. 2005). The mutant strain has a small decrease in Rubisco holoenzyme (Lim and Spreitzer, unpublished). It has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Phenotype: requires acetate, sensitive to light

Using plasmid p699 (GuhaMajumdar et al. 2008), Boon Hoe Lim in Spreitzer’s group performed rbcL direct mutagenesis, chloroplast transformation of wild-type 2137 mt+ (CC-4887), selection for spectinomycin resistance in the dark, and screening for a homoplasmic acetate-requiring phenotype to create 34 substitutions together in the Rubisco large subunit (A11V, V30E, V31T, R32K, M42V, C53A, D86H, I105L, V149Q, G168P, V221C, V235I, C256F, K258R, I265V, I282H, R305K, L326I, V341I, M349L, C369V, M375L, V391T, A398S, C399V, T428V, G442N, D443E, V444I, S447E, D470E, T471A, I472M, and K474T). The mutant lacks Rubisco holoenzyme (Lim and Spreitzer, unpublished). It was created to investigate phylogenetic differences that influence Rubisco catalysis (Du et al. 2003; Spreitzer et al. 2005). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or suppress light sensitivity (Spreitzer and Ogren 1983).


Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405

GuhaMajumdar M, Dawson-Baglien E, Sears BB (2008) Creation of a chloroplast microsatellite reporter for detection of replication slippage in Chlamydomonas reinhardtii. Eukaryot Cell 7:639-646

Spreitzer RJ, Ogren WL (1983) Nuclear suppressors of the photosensitivity associated with defective photosynthesis in Chlamydomonas reinhardtii. Plant Physiol 71:35-39

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created three substitutions (V221C, C256F, and K258R) together in the Rubisco large subunit. This mutant was created to investigate the molecular basis for the increased CO2/O2 specificity of mutant rbcL-Penta/rbcS1-ABSO mt+ (CC-4848) (Spreitzer et al. 2005). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), Boon Hoe Lim in Spreitzer’s group created two substitutions (V221C and I265V) together in the Rubisco large subunit. This mutant was created to investigate the molecular basis for the increased CO2/O2 specificity of mutant rbcL-Penta/rbcS1-ABSO mt+ (CC-4848) (Spreitzer et al. 2005). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL
  • Chromosome:
  • chloroplast

From Robert J. Spreitzer, University of Nebraska, November 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆/rbcS1-ABSO mt+ (Spreitzer et al. 2005), Boon Hoe Lim in Spreitzer’s group created two substitutions (C256F and K258R) together in the Rubisco large subunit. This mutant was created to investigate the molecular basis for the increased CO2/O2 specificity of mutant rbcL-Penta/rbcS1-ABSO mt+ (CC-4848) (Spreitzer et al. 2005). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.


Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230


  • Locus:
  • rbcL, RBCS1
  • Chromosome:
  • chloroplast,2

From Yingjun Wang and Martin Spalding, GDCB department, Iowa State University, December 2014

This LCIA (Nar1.2)-LCIB double mutant was generated in the ad1 mutant CC-4154 by insertional mutagenesis with aph8.


Wang, Spalding (2014) Acclimation to Very Low CO2: Contribution of Limiting CO2 Inducible Proteins, LCIB and LCIA, to Inorganic Carbon Uptake in Chlamydomonas reinhardtii. Plant Physiol 166:2040-2050


  • Locus:
  • LCIA [NAR1.2], LCIB
  • Chromosome:
  • 6,10

From Yingjun Wang and Martin Spalding, GDCB department, Iowa State University, December 2014

This is a walled progeny of CC-5064 lab1 mt- crossed with CC-620, carrying LCIA-LCIB double mutations.


Wang, Spalding (2014) Acclimation to Very Low CO2: Contribution of Limiting CO2 Inducible Proteins, LCIB and LCIA, to Inorganic Carbon Uptake in Chlamydomonas reinhardtii. Plant Physiol 166:2040-2050


  • Locus:
  • LCIA [NAR1.2], LCIB
  • Chromosome:
  • 6,10

From Yingjun Wang and Martin Spalding, GDCB department, Iowa State University, December 2014

This LCIA (Nar1.2) mutant is a progeny from a cross between CC-5064 lab1 mt- and CC-620.


Wang, Spalding (2014) Acclimation to Very Low CO2: Contribution of Limiting CO2 Inducible Proteins, LCIB and LCIA, to Inorganic Carbon Uptake in Chlamydomonas reinhardtii. Plant Physiol 166:2040-2050


  • Locus:
  • LCIA [NAR1.2]
  • Chromosome:
  • 6

From Yingjun Wang and Martin Spalding, GDCB department, Iowa State University, December 2014

This LCIA (Nar1.2) mutant is a progeny from a cross between CC-5064 lab1 mt- and CC-620.


Wang, Spalding (2014) Acclimation to Very Low CO2: Contribution of Limiting CO2 Inducible Proteins, LCIB and LCIA, to Inorganic Carbon Uptake in Chlamydomonas reinhardtii. Plant Physiol 166:2040-2050


  • Locus:
  • LCIA [NAR1.2]
  • Chromosome:
  • 6

From Janette Kropat, Merchant lab, UCLA, December 2014

Mutant created by transforming pArg-7.8 into CC-3960 arg7 crr1-2 mt+.


Castruita M, Casero D, Karpowicz SJ, Kropat J, Vieler A, Hsieh SI, Yan W, Cokus S, Loo JA, Benning C, Pellegrini M, Merchant SS (2011) Systems biology approach in Chlamydomonas reveals connections between copper nutrition and multiple metabolic steps. Plant Cell 23:1273-92

Hemschemeier A, Casero D, Liu B, Benning C, Pellegrini M, Happe T, Merchant SS (2013) Copper response regulator1-dependent and -independent responses of the Chlamydomonas reinhardtii transcriptome to dark anoxia. Plant Cell 25:3186-211

Sommer F, Kropat J, Malasarn D, Grossoehme NE, Chen X, Giedroc DP, Merchant SS (2010) The CRR1 nutritional copper sensor in Chlamydomonas contains two distinct metal-responsive domains. Plant Cell 22:4098-113


  • Locus:
  • CRR1
  • Chromosome:
  • 2

From Janette Kropat, Merchant lab, UCLA, December 2014

Mutant created by transforming pArg-7.8 into CC-3960 arg7 crr1-2 mt+.


Castruita M, Casero D, Karpowicz SJ, Kropat J, Vieler A, Hsieh SI, Yan W, Cokus S, Loo JA, Benning C, Pellegrini M, Merchant SS (2011) Systems biology approach in Chlamydomonas reveals connections between copper nutrition and multiple metabolic steps. Plant Cell 23:1273-92

Hemschemeier A, Casero D, Liu B, Benning C, Pellegrini M, Happe T, Merchant SS (2013) Copper response regulator1-dependent and -independent responses of the Chlamydomonas reinhardtii transcriptome to dark anoxia. Plant Cell 25:3186-211

Sommer F, Kropat J, Malasarn D, Grossoehme NE, Chen X, Giedroc DP, Merchant SS (2010) The CRR1 nutritional copper sensor in Chlamydomonas contains two distinct metal-responsive domains. Plant Cell 22:4098-113


  • Locus:
  • CRR1
  • Chromosome:
  • 2

From Janette Kropat, Merchant lab, UCLA, December 2014

crr1 mutant co-transformed with pArg-7.8 and WT CRR1.


Castruita M, Casero D, Karpowicz SJ, Kropat J, Vieler A, Hsieh SI, Yan W, Cokus S, Loo JA, Benning C, Pellegrini M, Merchant SS (2011) Systems biology approach in Chlamydomonas reveals connections between copper nutrition and multiple metabolic steps. Plant Cell 23:1273-92

Hemschemeier A, Casero D, Liu B, Benning C, Pellegrini M, Happe T, Merchant SS (2013) Copper response regulator1-dependent and -independent responses of the Chlamydomonas reinhardtii transcriptome to dark anoxia. Plant Cell 25:3186-211

Sommer F, Kropat J, Malasarn D, Grossoehme NE, Chen X, Giedroc DP, Merchant SS (2010) The CRR1 nutritional copper sensor in Chlamydomonas contains two distinct metal-responsive domains. Plant Cell 22:4098-113


  • Locus:
  • CRR1
  • Chromosome:
  • 2