Strains

From Bruce Kohorn, Duke University, January 1994

Phenotype: requires acetate; antibiotic resistant (spectinomycin, streptomycin)

This is a site-directed mutation transformed into the signal sequence of the petA gene. CC-125 was co-transformed with P-183 (carrying sr-u-2-60 spr-u-1-6-2 er-u-37) and a plasmid carrying a site-directed mutation in the signal sequence for the petA gene. Transformants were selected for spectinomycin and streptomycin resistance; the er-u-37 marker was not tested.

The A15E mutation introduces a change to GGAG at nucleotides 42-45 of the petA gene. This strain does not grow well on minimal medium.

Smith TA, Kohorn BD (1994) Mutations in a signal sequence for the thylakoid membrane identify multiple protein transport pathways and nuclear suppressors. J Cell Biol 126:365-374

• Locus:
• petA
• Chromosome:
• chloroplast

From Bruce Kohorn, Duke University, January 1994

Phenotype: requires acetate; antibiotic resistant (spectinomycin, streptomycin)

This is a site-directed mutation transformed into the signal sequence of the petA gene. CC-125 was co-transformed with P-183 (carrying sr-u-2-60 spr-u-1-6-2 er-u-37) and a plasmid carrying a site-directed mutation in the signal sequence for the petA gene. Transformants were selected for spectinomycin and streptomycin resistance; the er-u-37 marker was not tested.

The V16D mutation introduces a change to AGA at nucleotides 45-47 of the petA gene. This strain does not grow on minimal medium.

Smith TA, Kohorn BD (1994) Mutations in a signal sequence for the thylakoid membrane identify multiple protein transport pathways and nuclear suppressors. J Cell Biol 126:365-374

• Locus:
• petA
• Chromosome:
• chloroplast

From Bruce Kohorn, Duke University, January 1994

Phenotype: antibiotic resistant (spectinomycin, streptomycin)

This is a site-directed mutation transformed into the signal sequence of the petA gene. CC-125 was co-transformed with P-183 (carrying sr-u-2-60 spr-u-1-6-2 er-u-37) and a plasmid carrying a site-directed mutation in the signal sequence for the petA gene. Transformants were selected for spectinomycin and streptomycin resistance; the er-u-37 marker was not tested.

The M20E mutation introduces a change to GAGGCT at nucleotides 58-63 of the petA gene. This strain can grow on minimal medium.

Smith TA, Kohorn BD (1994) Mutations in a signal sequence for the thylakoid membrane identify multiple protein transport pathways and nuclear suppressors. J Cell Biol 126:365-374

• Locus:
• petA
• Chromosome:
• chloroplast

From Bruce Kohorn, Duke University, January 1994

Phenotype: antibiotic resistant (spectinomycin, streptomycin)

This is a site-directed mutation transformed into the signal sequence of the petA gene. CC-125 was co-transformed with P-183 (carrying sr-u-2-60 spr-u-1-6-2 er-u-37) and a plasmid carrying a site-directed mutation in the signal sequence for the petA gene. Transformants were selected for spectinomycin and streptomycin resistance; the er-u-37 marker was not tested.

The M20K mutation introduces a change to AGGCA at nucleotides 59-63 of the petA gene. This strain can grow on minimal medium.

Smith TA, Kohorn BD (1994) Mutations in a signal sequence for the thylakoid membrane identify multiple protein transport pathways and nuclear suppressors. J Cell Biol 126:365-374

• Locus:
• petA
• Chromosome:
• chloroplast

From Bruce Kohorn, Duke University, January 1994

Phenotype: requires acetate; antibiotic resistant (spectinomycin, streptomycin)

This is a site-directed mutation transformed into the signal sequence of the petA gene. CC-125 was co-transformed with P-183 (carrying sr-u-2-60 spr-u-1-6-2 er-u-37) and a plasmid carrying a site-directed mutation in the signal sequence for the petA gene. Transformants were selected for spectinomycin and streptomycin resistance; the er-u-37 marker was not tested.

The HCFS mutation produces a frame shift in the hydrophobic core by inserting a T after nucleotide 29 and removing nucleotide 45. This strain does not grow on minimal medium.

Smith TA, Kohorn BD (1994) Mutations in a signal sequence for the thylakoid membrane identify multiple protein transport pathways and nuclear suppressors. J Cell Biol 126:365-374

• Locus:
• petA
• Chromosome:
• chloroplast

From Bruce Kohorn, Duke University, January 1994

Phenotype: antibiotic resistant (spectinomycin, streptomycin)

This is a site-directed mutation transformed into the signal sequence of the petA gene. CC-125 was co-transformed with P-183 (carrying sr-u-2-60 spr-u-1-6-2 er-u-37) and a plasmid carrying a site-directed mutation in the signal sequence for the petA gene. Transformants were selected for spectinomycin and streptomycin resistance; the er-u-37 marker was not tested. The original transformant grew very slowly.

HC[delta] is a hydrophobic core deletion made by creating two EcoRI sites through sequential in vitro mutagenesis of nucleotides 39-43 and nucleotides 61-66, subcloning the 2.3 kb KpnI fragment containing this modified petA gene into a pUC119 plasmid lacking both HindIII and EcoRI sites, digesting with EcoRI to remove the hydrophobic core, and then religating.

Smith TA, Kohorn BD (1994) Mutations in a signal sequence for the thylakoid membrane identify multiple protein transport pathways and nuclear suppressors. J Cell Biol 126:365-374

• Locus:
• petA
• Chromosome:
• chloroplast

From Peter Heifetz, Boynton-Gillham laboratory, Duke University, April 1994

Phenotype: can grow at very high light intensity

This strain, which grows well at very high light intensities, was originally described as “125 S-1 high light suppressor”. This is a spontaneous mutant selected, as described by Förster et al. (1999), in the wild-type strain CC-125 for ability to grow and remain green at very high light intensity (1500-2000 micromole photons per square meter per second).

Förster B, Osmond B, Boynton JE, Gillham NW (1999) Mutants of Chlamydomonas reinhardtii resistant to very high light. J Photochem Photobiol 48:127-135

From Peter Heifetz, Boynton-Gillham laboratory, Duke University, April 1994

Phenotype: can grow at very high light intensity

This strain, which grows well at very high light intensities, was originally described as “125 S-6 high light suppressor”. This is a spontaneous mutant selected, as described by Förster et al. (1999), in the wild-type strain CC-125 for ability to grow and remain green at very high light intensity (1500-2000 micromole photons per square meter per second).

Förster B, Osmond B, Boynton JE, Gillham NW (1999) Mutants of Chlamydomonas reinhardtii resistant to very high light. J Photochem Photobiol 48:127-135

From Gary Small, University of South Dakota, April 1994

Phenotype: UV sensitive

This strain is UV-sensitive and deficient in photoreactivation. It was isolated by Cox and Small in the 137c background, after mutagenesis with nitrosoguanidine.

Cox JL, Small GD (1985) Isolation of a photoreactivation-deficient mutant of Chlamydomonas. Mutat Res 146:249-255

Small GD (1987) Repair systems for nuclear and chloroplast DNA in Chlamydomonas reinhardtii. Mut Res 181:31–35

Miadoková E, Podstavková S, Vlček D (1991) Induction of new uv-sensitive mutants of chlamydomonas reinhardtii. Archiv für Protistenkunde 139:207-212

Vlček D, Podstavková S, Miadoková E (1995) Interactions between photolyase and dark repair processes in Chlamydomonas reinhardtii. Mutat Res 336:251-256

Petersen JL, Ronan PJ (2010) Critical role of 7,8-didemethyl-8-hydroxy-5-deazariboflavin for photoreactivation in Chlamydomonas reinhardtii. J Biol Chem 285:32467-32475

• Locus:
• PHR1
• Chromosome:
• 17

From Jonathan Jarvik, Carnegie-Mellon University, July 1994

Phenotype: impaired motility

This mutant was originally designated cs89 and was described by Jarvik et al. in a presentation at the 1976 meeting of the American Society for Cell Biology as “a cold-sensitive mutant with aberrant control of flagellar length.” Barsel et al. determined that this was an lf2 allele.

Please see CC-803 for more information on the LF2 locus.

Jarvik J, Lefebvre PA, Rosenbaum JL (1976) A cold-sensitive mutant of Chlamydomonas reinhardi with aberrant control of flagellar length. J Cell Biol 70:140a

Barsel SE, Wexler DE, Lefebvre PA (1988) Genetic analysis of long-flagella mutants of Chlamydomonas reinhardtii. Genetics 118:637-648

• Locus:
• LF2
• Chromosome:
• 12

From Carolyn Silflow, University of Minnesota, July 1994

Phenotype: herbicide resistant (amiprophos methyl, oryzalin)

This is a single colony isolate of the strain SJ863-D6 (tua1 nit2 mt-). The original mutant was designated upA12. The tua1 mutation is a missense mutation changing Tyr to Hys at amino acid residue 24 in the alpha-tubulin 1 gene. It confers resistance to 8.5 micromolar amiprophos methyl. The tua1 mutant is also resistant to oryzalin, and is more sensitive to taxol than wild type.

James SW, Silflow CD, Stroom P, Lefebvre PA (1993) A mutation in the α-tubulin gene of Chlamydomonas reinhardtii confers resistance to anti-microtubule herbicides. J Cell Sci 106:209-218

• Locus:
• NIT2, TUA1
• Chromosome:
• 3

From Robert Togasaki, Indiana University, July 1994

From Robert Togasaki, Indiana University, July 1994, originally from Masa and Kaoru Kitayama

From Robert Togasaki, Indiana University, July 1994, originally from Masa and Kaoru Kitayama

From Jon Suzuki, Nagoya University September 1994

Phenotype: antibiotic resistant (spectinomycin, streptomycin)

This is a transformant of strain 2137 (see CC-3269) with 16S rRNA mutations to streptomycin and spectinomycin resistance (carried by plasmid P-183). This was used as a control strain for the chlL insertional mutant JS1000 (see CC-3840).

Suzuki JY, Bauer CE (1992) Light-independent chlorophyll biosynthesis: involvement of the chloroplast gene chlL (frxC). Plant Cell 4:929-940

From Krishna Niyogi, Carnegie Institution of Washington, Stanford, September 1994

Phenotype: requires arginine

This is Niyogi’s strain KNX31-2. It was derived from a cross of CC-2678 nit1-305 cw15 x CC-424 cw15 arg2, followed by backcrosses to wild type (CC-124 and CC-125). It has the wild type alleles at the NIT2 (inferred because it can grow on nitrite medium) and CW15 loci. The nit1 allele could be either nit1-305 from CC-2678 or nit1-137, which is carried by CC-424, CC-124, and CC-125. (Statistically the latter is more likely, but it hasn’t been tested).

For more information on the arg7-8 (arg2) mutation, please see CC-48.

• Locus:
• ARG7, NIT1
• Chromosome:
• 1,9

From George Witman, Worcester Foundation for Experimental Biology, October 1994

Phenotype: altered phototaxis

This is an undocumented mutant that probably has negative phototaxis.

From George Witman, Worcester Foundation for Experimental Biology, October 1994

Phenotype: altered phototaxis

This is an undocumented mutant that probably has negative phototaxis.

From George Witman, Worcester Foundation for Experimental Biology, October 1994

Phenotype: impaired motility

This is a mutant with the mbo (“moves backwards only”) phenotype. We don’t know if it’s an allele at any of the named MBO loci.

Nakamura S (1979) A backward swimming mutant of Chlamydomonas reinhardii. Exp Cell Res 123:441-444

Nakamura S (1981) Two Different Backward-Swimming Mutants of Chlamydomonas reinhardtii. Cell Struct Funct 6:385-393

From George Witman, Worcester Foundation for Experimental Biology, October 1994

Phenotype: impaired motility

This is a mutant with the mbo (“moves backwards only”) phenotype. We don’t know if it’s an allele at any of the named MBO loci.

Nakamura S (1981) Two Different Backward-Swimming Mutants of Chlamydomonas reinhardtii. Cell Struct Funct 6:385-393

From George Witman, Worcester Foundation for Experimental Biology, October 1994

Phenotype: uniflagellate

This is a uniflagellate mutant isolated by Kamiya. In a letter to Witman, Kamiya reported that like other uni mutants, uniflagellate cells of uf401 lack the cis-flagellum, and that cultures of this strain contain a mixture of uniflagellate and flagella-less cells. A count of 300 cells of this strain showed 55% uniflagellate, 4% biflagellate, and 41% flagella-less cells.

From George Witman, Worcester Foundation for Experimental Biology, October 1994

This strain, originally obtained by Witman from Luck, is “a descendant of an extragenic suppressor of pf6 obtained with diepoxybutane as a mutagen….D22.2A showed co-segregation of the suppressor motility phenotype and the altered mobility of the beta heavy chain on a 3.2% acrylamide 6M urea 1-D gel.”

From George Witman, Worcester Foundation for Experimental Biology, October 1994

Phenotype: impaired motility

This strain was sent to Witman by Philip Filner of Michigan State University, and was described as follows: “normal length flagella that function, but cells spin rather than move linearly and some cells have 3-5 flagella instead of just 2.”

From George Witman, Worcester Foundation for Experimental Biology, October 1994

Phenotype: impaired motility

This strain was sent to Witman by Philip Filner of Michigan State University, and was described as follows: “normal length flagella that beat slowly, so cells swim slowly. (This is not the original phenotype, but the cells nevertheless appear to be mutant.)”

From George Witman, Worcester Foundation for Experimental Biology, October 1994

Phenotype: impaired motility

This strain was sent to Witman by Philip Filner of Michigan State University, and was described as follows: “normal length flagella that twitch. Cells do not swim, i.e. are semi-paralyzed.”

From George Witman, Worcester Foundation for Experimental Biology, October 1994

Phenotype: impaired motility

This mutant has the alpha and gamma outer arm dynein heavy chains, and a novel peptide of ca. 160 kDa that appears to be a truncated beta chain.

Sakakibara H, Takada S, King SM, Witman GB, Kamiya R (1993) A Chlamydomonas outer arm dynein mutant with a truncated beta heavy chain. J Cell Biol 122:653-661

• Locus:
• ODA4 [DHC14]
• Chromosome:
• 9

From George Witman, Worcester Foundation for Experimental Biology, October 1994; originally from David Luck, Rockefeller University

This is the pf14 mutant (but which allele is uncertain) with the suppressor mutation sup-pf3. Please see CC-613 and CC-1399 for more information on these mutants.

• Locus:
• PF14 [RSP3], SUP-PF3 [PF2]
• Chromosome:
• 6,11

From George Witman, Worcester Foundation for Experimental Biology, October 1994; originally from David Luck, Rockefeller University

This is the pf1 mutant (see CC-1024) with the suppressor mutation sup-pf3.

• Locus:
• PF1 [RSP4], SUP-PF3 [PF2]
• Chromosome:
• 5,6,11

From George Witman, Worcester Foundation for Experimental Biology, October 1994; originally from P.A. Lefebvre, University of Minnesota,

Phenotype: impaired motility; temperature sensitive

The tnr1 mutant was isolated by Larkin et al. in strain NO 3C mt- [a wild-type segregant from a cross of 137C derivative lines equivalent to CC-621]. Cells with the tnr1 mutation cannot regenerate flagella at 34 degrees, and die after 4-5 days at this temperature.

Larkin JC, Lefebvre PA, Silflow CD (1989) A gene essential for viability and flagellar regeneration maps to the uni linkage group of Chlamydomonas reinhardtii. Curr Genet 15:377-384

• Locus:
• TNR1
• Chromosome:
• 17

From George Witman, Worcester Foundation for Experimental Biology, October 1994; originally from P.A. Lefebvre, University of Minnesota

Phenotype: impaired motility; temperature sensitive

Please see CC-3002 for more information on tnr1.

• Locus:
• TNR1
• Chromosome:
• 17