Strains

From Arminio Boschetti, University of Bern, August 2001

Phenotype: chlorophyll and carotenoid deficient; requires arginine

From Boschetti’s notes on this strain: The pg-113 mutant was isolated by M. Tellenbach in Boschetti’s laboratory, by selecting cells with altered color after UV mutagenesis of an arg2 strain (CCAP 11/32f, Levine arg2 mt+) and subsequently determined in collaboration with Alexander Chunayev to be an allele at the CBN1 locus although its phenotype differs somewhat from the cbn1-48 mutant that defines that locus. It lacks a stable chlorophyll complex and chlorophyll b, but has normal chlorophyll a., and the LHCII proteins are normal. The amount of neoxanthin is reduced. This strain grows phototrophically but should be kept at low to medium light intensity.

Michel H, Tellenbach M, Boschetti A (1983) A chlorophyll b-less mutant of Chlamydomonas reinhardii lacking in the light-harvesting chlorophyll complex but not in its apoproteins. Biochim Biophys Acta 725:417-424

Hoober JK, Maloney MA, Asbury LR, Marks DB (1990) Accumulation of Chlorophyll a/b-Binding Polypeptides in Chlamydomonas reinhardtii y-1 in the Light or Dark at 38 degrees C : Evidence for Proteolytic Control. Plant Physiol 92:419-426

Hoober JK, Hughes MJ (1992) Purification and Characterization of a Membrane-Bound Protease from Chlamydomonas reinhardtii. Plant Physiol 99:932-937

• Locus:
• PG-113 [CAO1]
• Chromosome:
• 1

From Arminio Boschetti, University of Bern, August 2001

Phenotype: carotenoid deficient; requires arginine

The pg-101 or lor1 mutant was isolated by M. Tellenbach in Boschetti’s laboratory, by selecting cells with altered color after UV mutagenesis of an arg2 strain (CCAP 11/32f, Levine arg2 mt+). Tellenbach characterized it as deficient in loroxanthin. Subsequently Chunayev et al. showed that it was deficient in alpha cyclase, resulting in failure to produce the carotenoid epsilon rings.

Please see CC-2420 for additional information on this mutant.

This strain should be kept in dim light.

Chunayev AS, Mirnaya ON, Maslov VG, Boschetti A (1991) Chlorophyll b and chloroxanthin-deficient mutants of Chlamydomonas reinhardtii. Photosynthetica 25:291-301

From Arminio Boschetti, University of Bern, August 2001, his 619f

Phenotype: requires acetate; wall deficient

F54 is a nuclear gene mutation that affects translation of the chloroplast atpA mRNA. This strain was obtained by crossing CC-980 F54 mt+ x CC-1883 cw15 mt.

Boschetti A, Schmid K (1998) Energy Supply for ATP-Synthase Deficient Chloroplasts of Chlamydomonas reinhardii. Plant Cell Physiol 39:160-168

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

CC-3875 was created by transformation of T60-3 (rbcS-T60-3T60-3) with a modified plasmid pRbcS-2. The original plasmid, a pUC19-vector containing the genomic BamHI/EcoRI fragment encoding rbcS-2 [EMBL/GenBank accession X04471], total length 10.5 kb, was shortened by elimination of an unknown DNA sequence of about 5 kb. Transformation resulted in complementation of T60-3 to normal photoautotrophy. CC-3875 was used as a control strain for CC-3876 through CC-3889, which have mutations in the RBCS transit peptide.

Khrebtukova I, Spreitzer RJ (1996) Elimination of the Chlamydomonas gene family that encodes the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase. Proc Natl Acad Sci USA 93:13689-13693

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3876, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 1 in the transit peptide of the precursor of the Rubisco small subunit such that Pro(24)/Met(25) are exchanged by Ile(24)/Trp(25) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to normal photoautotrophy. The intermediate sized small subunit is no longer produced.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3877, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 1 in the transit peptide of the precursor of the Rubisco small subunit such that Pro(24)/Met(25) are exchanged by Ile(24)/Trp(25) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to normal photoautotrophy. The intermediate sized small subunit is no longer produced.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3878, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 1 in the transit peptide of the precursor of the Rubisco small subunit such that Pro(24)/Met(25) are exchanged by Ile(24)/Trp(25) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to normal photoautotrophy. The intermediate sized small subunit is no longer produced.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3880, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 1 in the transit peptide of the precursor of the Rubisco small subunit such that Pro(24)/Met(25) are exchanged by Ile(24)/Trp(25) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to normal photoautotrophy. The intermediate sized small subunit is no longer produced.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3882, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 2 in the transit peptide of the precursor of the Rubisco small subunit such that Met(46) is exchanged by His(46) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to normal photoautotrophy. Synthesis of intermediate sized small subunit is enhanced, but mature small subunit is also formed.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3883, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 2 in the transit peptide of the precursor of the Rubisco small subunit such that Met(46) is exchanged by His(46) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to normal photoautotrophy. Synthesis of intermediate sized small subunit is enhanced, but mature small subunit is also formed.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3886, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 2 in the transit peptide of the precursor of the Rubisco small subunit such that Gln(45)/Met(46) are exchanged by Ile(45)/His(46) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to photoautotrophy, but strains grow more slowly. Formation of intermediate sized small subunit is very much enhanced. No mature small subunit was detected by Western blotting, but some immuno cross-reacting proteins of slightly higher molecular masses were detected.”

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3887, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 2 in the transit peptide of the precursor of the Rubisco small subunit such that Gln(45)/Met(46) are exchanged by Ile(45)/His(46) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to photoautotrophy, but strains grow more slowly. Formation of intermediate sized small subunit is very much enhanced. No mature small subunit was detected by Western blotting, but some immuno cross-reacting proteins of slightly higher molecular masses were detected.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3888, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 2 in the transit peptide of the precursor of the Rubisco small subunit such that Gln(45)/Met(46) are exchanged by Ile(45)/His(46) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to photoautotrophy, but strains grow more slowly. Formation of intermediate sized small subunit is very much enhanced. No mature small subunit was detected by Western blotting, but some immuno cross-reacting proteins of slightly higher molecular masses were detected.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3889, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 2 in the transit peptide of the precursor of the Rubisco small subunit such that Gln(45)/Met(46) are exchanged by Ile(45)/His(46) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to photoautotrophy, but strains grow more slowly. Formation of intermediate sized small subunit is very much enhanced. No mature small subunit was detected by Western blotting, but some immuno cross-reacting proteins of slightly higher molecular masses were detected.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3890, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 2 in the transit peptide of the precursor of the Rubisco small subunit such that Gln(45)/Met(46) are exchanged by Ile(45)/His(46) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to photoautotrophy, but strains grow more slowly. Formation of intermediate sized small subunit is very much enhanced. No mature small subunit was detected by Western blotting, but some immuno cross-reacting proteins of slightly higher molecular masses were detected.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3891, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 2 in the transit peptide of the precursor of the Rubisco small subunit such that Gln(45)/Met(46) are exchanged by Ile(45)/His(46) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to photoautotrophy, but strains grow more slowly. Formation of intermediate sized small subunit is very much enhanced. No mature small subunit was detected by Western blotting, but some immuno cross-reacting proteins of slightly higher molecular masses were detected.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3892, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 2 in the transit peptide of the precursor of the Rubisco small subunit such that Gln(45)/Met(46) are exchanged by Ile(45)/His(46) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to photoautotrophy, but strains grow more slowly. Formation of intermediate sized small subunit is very much enhanced. No mature small subunit was detected by Western blotting, but some immuno cross-reacting proteins of slightly higher molecular masses were detected.

• Locus:
• RBCS2
• Chromosome:

From Arminio Boschetti, University of Bern, August 2001

CC-3875 through CC-3893 are a collection of transformants of the T60-3 Rubisco knockout mutant created by Khrebtukova and Spreitzer (CC-4415).

For CC-3893, the plasmid used for the transformation leading to CC-3875 was modified by mutating processing site 2 in the transit peptide of the precursor of the Rubisco small subunit such that Gln(45)/Met(46) are exchanged by Ile(45)/His(46) and a NdeI restriction site is introduced into the DNA. Transformation resulted in complementation of rbcS-T60-3 to photoautotrophy, but strains grow more slowly. Formation of intermediate sized small subunit is very much enhanced. No mature small subunit was detected by Western blotting, but some immuno cross-reacting proteins of slightly higher molecular masses were detected.

• Locus:
• RBCS2
• Chromosome:

From Mary Porter, University of Minnesota, October 2001

Phenotype: impaired motility

This is a mt+ isolate of pf16B. Please see CC-1034 for more information on the PF16 locus, and CC-2510 for pf16B.

Porter ME, Power J, Dutcher SK (1992) Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol 118:1163-1176

• Locus:
• PF16
• Chromosome:
• 9

From Mary Porter, University of Minnesota, October 2001

Phenotype: impaired motility

This is a mt- isolate of pf16B. Please see CC-1034 for more information on the PF16 locus, and CC-2510 for pf16B.

Porter ME, Power J, Dutcher SK (1992) Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol 118:1163-1176

• Locus:
• PF16
• Chromosome:
• 9

From Mary Porter, University of Minnesota, October 2001

pf16BR3 is an intragenic revertant of pf16B. Please see CC-1034 for more information on the PF16 locus, CC-2510 for pf16B, and CC-2511 for pf16B-R3.

Porter ME, Power J, Dutcher SK (1992) Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol 118:1163-1176

• Locus:
• PF16
• Chromosome:
• 9

From Mary Porter, University of Minnesota, October 2001

The pf9-2 mutation was isolated as a suppressor of the temperature-sensitive mutant pf16BR3 (please see CC-2511), which restores motility at 32 degrees. In the absence of the pf16BR3 mutation, pf9-2 cells swim more slowly than wild type.

Porter ME, Power J, Dutcher SK (1992) Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol 118:1163-1176

• Locus:
• PF9 [DHC1], PF16
• Chromosome:
• 9

From Mary Porter, University of Minnesota, October 2001

This is a pf9 allele isolated as a bypass suppressor of pf16BR3 (please see CC-3897). For more information on the PF9 locus, please see CC-1254.

Porter ME, Power J, Dutcher SK (1992) Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol 118:1163-1176

• Locus:
• PF9 [DHC1]
• Chromosome:
• 12

From Mary Porter, University of Minnesota, October 2001

This is a pf9 allele isolated as a bypass suppressor of pf16BR3 (please see CC-3897). For more information on the PF9 locus, please see CC-1254.

Porter ME, Power J, Dutcher SK (1992) Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol 118:1163-1176

• Locus:
• PF9 [DHC1]
• Chromosome:
• 12

From Mary Porter, University of Minnesota, October 2001

Phenotype: impaired motility

This is a double mutant affecting the outer and inner dynein arms. The flagella are short and paralyzed.

For more information on the pf9-2 mutation, please see CC-3897, and for pf28, please see CC-1877.

Porter ME, Power J, Dutcher SK (1992) Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol 118:1163-1176

Myster SH, Knott JA, Wysocki KM, O'Toole E, Porter ME (1999) Domains in the 1alpha dynein heavy chain required for inner arm assembly and flagellar motility in Chlamydomonas. J Cell Biol 146:801-818

• Locus:
• PF9 [DHC1], PF28 [DHC15]
• Chromosome:
• 11,12

From Mary Porter, University of Minnesota, October 2001

Phenotype: impaired motility

This is a double mutant affecting the outer and inner dynein arms, with a third extragenic suppressor mutation that partially restores I1 inner arm function.

For more information on the pf9-2 mutation, please see CC-3897 and for pf28, please see CC-1877.

Porter ME, Power J, Dutcher SK (1992) Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol 118:1163-1176

Myster SH, Knott JA, Wysocki KM, O'Toole E, Porter ME (1999) Domains in the 1alpha dynein heavy chain required for inner arm assembly and flagellar motility in Chlamydomonas. J Cell Biol 146:801-818

• Locus:
• PF9 [DHC1], PF28 [DHC15]
• Chromosome:
• 11,12

From Mary Porter, University of Minnesota, October 2001

Phenotype: impaired motility

This is a double mutant affecting the outer and inner dynein arms, with a third extragenic suppressor mutation that partially restores I1 inner arm function.

For more information on the pf9-2 mutation, please see CC-3897 and for pf28, please see CC-1877.

Porter ME, Power J, Dutcher SK (1992) Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol 118:1163-1176

Myster SH, Knott JA, Wysocki KM, O'Toole E, Porter ME (1999) Domains in the 1alpha dynein heavy chain required for inner arm assembly and flagellar motility in Chlamydomonas. J Cell Biol 146:801-818

• Locus:
• PF9 [DHC1], PF28 [DHC15]
• Chromosome:
• 11,12

From Mary Porter, University of Minnesota, October 2001

Phenotype: impaired motility

This is a double mutant affecting the outer and inner dynein arms, with a third extragenic suppressor mutation that restores wild type flagellar length without restoring outer arm or I1 inner arm function.

For more information on the pf9-2 mutation, please see CC-3897 and for pf28, please see CC-1877.

Porter ME, Power J, Dutcher SK (1992) Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol 118:1163-1176

Myster SH, Knott JA, Wysocki KM, O'Toole E, Porter ME (1999) Domains in the 1alpha dynein heavy chain required for inner arm assembly and flagellar motility in Chlamydomonas. J Cell Biol 146:801-818

• Locus:
• PF9 [DHC1], PF28 [DHC15]
• Chromosome:
• 11,12

From Mary Porter, University of Minnesota, October 2001

For more information on the pf9-2 mutation, please see CC-3897, and for sup-pf1 please see CC-1397.

Porter ME, Power J, Dutcher SK (1992) Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol 118:1163-1176

Myster SH, Knott JA, Wysocki KM, O'Toole E, Porter ME (1999) Domains in the 1alpha dynein heavy chain required for inner arm assembly and flagellar motility in Chlamydomonas. J Cell Biol 146:801-818

• Locus:
• PF9 [DHC1], SUP-PF1 [DHC14]
• Chromosome:
• 9,12

From Mary Porter, University of Minnesota, October 2001

Phenotype: antibiotic resistant (streptomycin)

This strain carries the original sup-pf1 allele (please see CC-1397), and was used by Porter et al. for RFLP analysis with C. smithii.

Porter ME, Knott JA, Gardner LC, Mitchell DR, Dutcher SK (1994) Mutations in the SUP-PF-1 locus of Chlamydomonas reinhardtii identify a regulatory domain in the beta-dynein heavy chain. J Cell Biol 126:1495-1507

• Locus:
• SR1, SUP-PF1 [DHC14]
• Chromosome:
• 9