Strains

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

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

Using established methods of directed mutagenesis and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) (Genkov and Spreitzer 2009), Todor Genkov in Spreitzer’s group created a Y32D substitution in the Rubisco small subunit, which causes a decrease in Rubisco CO2/O2 specificity and holoenzyme stability (Genkov and Spreitzer, unpublished). This mutant can grow on minimal medium at 25 °C, but dies on minimal medium at 35 °C. The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

Genkov T, Spreitzer RJ (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J Biol Chem 284:30105-30112

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

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

Using established methods of directed mutagenesis and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) (Genkov and Spreitzer 2009), Todor Genkov in Spreitzer’s group created a Y32E substitution in the Rubisco small subunit, which causes a decrease in Rubisco CO2/O2 specificity and holoenzyme stability (Genkov and Spreitzer, unpublished). This mutant can grow on minimal medium at 25 °C, but dies on minimal medium at 35 °C. The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

Genkov T, Spreitzer RJ (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J Biol Chem 284:30105-30112

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

Using established methods of directed mutagenesis and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) (Genkov and Spreitzer 2009), Todor Genkov in Spreitzer’s group created a Y32F substitution in the Rubisco small subunit (Genkov and Spreitzer, unpublished). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

Genkov T, Spreitzer RJ (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J Biol Chem 284:30105-30112

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

Using established methods of directed mutagenesis and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) (Genkov and Spreitzer 2009), Todor Genkov in Spreitzer’s group created a Y32R substitution in the Rubisco small subunit, which causes a decrease in Rubisco holoenzyme stability (Genkov and Spreitzer, unpublished). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

Genkov T, Spreitzer RJ (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J Biol Chem 284:30105-30112

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

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

Directed mutagenesis and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) were used to create an E43A substitution (GAG-GCC) in the Rubisco small subunit, which causes a decrease in Rubisco CO2/O2 specificity and holoenzyme stability (Genkov and Spreitzer 2009). This mutant can grow on minimal medium at 25 °C, but dies on minimal medium at 35 °C. The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

Genkov T, Spreitzer RJ (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J Biol Chem 284:30105-30112

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

Using a plasmid that would create an E43A substitution (GAG-GCC) in the Rubisco small subunit, which causes a decrease in Rubisco CO2/O2 specificity and holoenzyme stability (Genkov and Spreitzer 2009), Todor Genkov in Spreitzer’s group transformed rbcS∆-CAL005.01.13 pf2 mt+ (CC-4692). In contrast to the E43A mutant described previously (Genkov and Spreitzer 2009), this strain has a cell wall and can be used in genetic crosses. This mutant can grow on minimal medium at 25 °C, but dies on minimal medium at 35 °C. The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

Genkov T, Spreitzer RJ (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J Biol Chem 284:30105-30112

  • Locus:
  • RBCS1, PF2
  • Chromosome:
  • 2,11

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

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

Directed mutagenesis and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) were used to create a W73A substitution (TGG-GCC) in the Rubisco small subunit, which causes a decrease in Rubisco carboxylation catalytic efficiency and holoenzyme stability (Genkov and Spreitzer 2009). This mutant can grow on minimal medium at 25 °C, but dies on minimal medium at 35 °C. The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

Genkov T, Spreitzer RJ (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J Biol Chem 284:30105-30112

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

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

Directed mutagenesis and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) were used to create an L78A substitution (CTG-GCC) in the Rubisco small subunit, which causes a decrease in Rubisco carboxylation catalytic efficiency and holoenzyme stability (Genkov and Spreitzer 2009). This mutant can grow on minimal medium at 25 °C, but dies on minimal medium at 35 °C. The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability

Genkov T, Spreitzer RJ (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J Biol Chem 284:30105-30112

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

Directed mutagenesis and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) were used to create a P79A substitution (CCC-GCC) in the Rubisco small subunit, which causes a decrease in Rubisco carboxylation catalytic efficiency and holoenzyme stability (Genkov and Spreitzer 2009). The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

Genkov T, Spreitzer RJ (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J Biol Chem 284:30105-30112

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

Directed mutagenesis and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) were used to create an F81A substitution (TTC-GCC) in the Rubisco small subunit, which causes a decrease in Rubisco carboxylation catalytic efficiency and holoenzyme stability (Genkov and Spreitzer 2009). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

Genkov T, Spreitzer RJ (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J Biol Chem 284:30105-30112

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

Genetic engineering and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) were used to remove intron 3 from the coding region of the Rubisco small subunit (Genkov et al. 2010). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

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

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

Genetic engineering and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) were used to move introns 1 and 2 into the transit-peptide coding region of the Rubisco small subunit (Genkov et al. 2010). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

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

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

Phenotype: altered CO2 assimilation

Genetic engineering and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) were used to create a hybrid Rubisco enzyme comprised of Chlamydomonas large subunits and spinach small subunits (Genkov et al. 2010). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

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

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

Phenotype: altered CO2 assimilation

Using a plasmid that encodes the spinach-Rubisco small subunit (Genkov et al. 2010), Todor Genkov in Spreitzer’s group transformed rbcS∆-CAL005.01.13 pf2 mt+ (CC-4692). In contrast to the SSSO mutant described previously (Genkov et al. 2010), this strain has a cell wall and can be used in genetic crosses. The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

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

  • Locus:
  • RBCS1, PF2
  • Chromosome:
  • 2,11

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

Phenotype: altered CO2 assimilation

Genetic engineering and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) were used to create a hybrid Rubisco enzyme comprised of Chlamydomonas large subunits and Arabidopsis small subunits (Genkov et al. 2010). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

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

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

Phenotype: altered CO2 assimilation

Using a plasmid that encodes the Arabidopsis-Rubisco small subunit (Genkov et al. 2010), Todor Genkov in Spreitzer’s group transformed rbcS∆-CAL005.01.13 pf2 mt+ (CC-4692). In contrast to the SSAT mutant described previously (Genkov et al. 2010), this strain has a cell wall and can be used in genetic crosses. The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

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

  • Locus:
  • RBCS1, PF2
  • Chromosome:
  • 2,11

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

Phenotype: altered CO2 assimilation

Genetic engineering and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) were used to create a hybrid Rubisco enzyme comprised of Chlamydomonas large subunits and sunflower small subunits (Genkov et al. 2010). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

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

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

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

Phenotype: altered CO2 assimilation

Using a plasmid that encodes the sunflower-Rubisco small subunit (Genkov et al. 2010), Todor Genkov in Spreitzer’s group transformed rbcS∆-CAL005.01.13 pf2 mt+ (CC-4692). In contrast to the SSHA mutant described previously (Genkov et al. 2010), this strain has a cell wall and can be used in genetic crosses. The strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

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

  • Locus:
  • RBCS1, PF2
  • Chromosome:
  • 2,11

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

Phenotype: altered CO2 assimilation

Genetic engineering and nuclear-gene transformation of rbcS∆-T60-3 mt- (CC-4415) (Genkov et al. 2010) were used to create a hybrid Rubisco enzyme comprised of Chlamydomonas large subunits and tobacco small subunits for studies of Rubisco activase (Wachter et al. 2013). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function or stability.

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

Wachter RM, Salvucci ME, Carmo-Silva AE, Barta C, Genkov T, Spreitzer RJ (2013) Activation of interspecies-hybrid Rubisco enzymes to assess different models for the Rubisco-Rubisco activase interaction. Photosynth Res 117:557-566

  • Locus:
  • RBCS1
  • Chromosome:
  • 2

From Lai-Wa Tam, Lefebvre lab, University of Minnesota, July 2014

Phenotype: very long flagella

Tam LW, Ranum PT, Lefebvre PA (2013) CDKL5 regulates flagellar length and localizes to the base of the flagella in Chlamydomonas. Mol Biol Cell 24:588-600

  • Locus:
  • LF4
  • Chromosome:
  • 13

From Lai-Wa Tam, Lefebvre lab, University of Minnesota, July 2014

Phenotype: unequal length flagella

Tam LW, Ranum PT, Lefebvre PA (2013) CDKL5 regulates flagellar length and localizes to the base of the flagella in Chlamydomonas. Mol Biol Cell 24:588-600

  • Locus:
  • ARG7
  • Chromosome:
  • 1

From Lai-Wa Tam, Lefebvre lab, University of Minnesota, July 2014

5′ region of LF1 deleted, can be rescued by transformation. Isolated by Carolyn Silfow, U of MN.

  • Locus:
  • LF1
  • Chromosome:
  • 2

From Lai-Wa Tam, Lefebvre lab, University of Minnesota, July 2014

Phenotype: normal length flagella, slow swimmer

Rescue of lf1-1 by HA-tagged LF1

Nguyen RL, Tam LW, Lefebvre PA (2005) The LF1 gene of Chlamydomonas reinhardtii encodes a novel protein required for flagellar length control. Genetics 169:1415-24

  • Locus:
  • LF1
  • Chromosome:
  • 2

From Lai-Wa Tam, Lefebvre lab, University of Minnesota, July 2014

Phenotype: normal length flagella, slow swimmer

Rescue of lf1-1 by HA-tagged LF1

Nguyen RL, Tam LW, Lefebvre PA (2005) The LF1 gene of Chlamydomonas reinhardtii encodes a novel protein required for flagellar length control. Genetics 169:1415-24

  • Locus:
  • LF1
  • Chromosome:
  • 2

From Lai-Wa Tam, Lefebvre lab, University of Minnesota, July 2014

Rescue of lf3-5 with HA-tagged LF3

Tam LW, Dentler WL, Lefebvre PA (2003) Defective flagellar assembly and length regulation in LF3 null mutants in Chlamydomonas. J Cell Biol 163:597-607

  • Locus:
  • LF3
  • Chromosome:
  • 1

From Lai-Wa Tam, Lefebvre lab, University of Minnesota, July 2014

Rescue of lf3-5 with HA-tagged LF3

Tam LW, Dentler WL, Lefebvre PA (2003) Defective flagellar assembly and length regulation in LF3 null mutants in Chlamydomonas. J Cell Biol 163:597-607

  • Locus:
  • LF3
  • Chromosome:
  • 1

From Lai-Wa Tam, Lefebvre lab, University of Minnesota, July 2014

Phenotype: long flagella

Tam LW, Dentler WL, Lefebvre PA (2003) Defective flagellar assembly and length regulation in LF3 null mutants in Chlamydomonas. J Cell Biol 163:597-607

  • Locus:
  • LF2 [CDKI1]
  • Chromosome:
  • 12

From Lai-Wa Tam, Lefebvre lab, University of Minnesota, July 2014

Phenotype: long flagella

Tam LW, Dentler WL, Lefebvre PA (2003) Defective flagellar assembly and length regulation in LF3 null mutants in Chlamydomonas. J Cell Biol 163:597-607

  • Locus:
  • LF2 [CDKI1]
  • Chromosome:
  • 12

From Lai-Wa Tam, Lefebvre lab, University of Minnesota, July 2014

Phenotype: mostly flagella-less

  • Locus:
  • LF1, LF2 [CDKI1]
  • Chromosome:
  • 2,12

From Lai-Wa Tam, Lefebvre lab, University of Minnesota, July 2014

Phenotype: mostly flagella-less

  • Locus:
  • LF1, LF2 [CDKI1]
  • Chromosome:
  • 2,12