Strains
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: HAP2-R185Q in 40D4
Background strain: 40D4
Plasmid transformation: HAP2-R185Q-aphVIII plasmid transformed into 40D4, one plasmid
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: MAR1-Flag knock-down cell, point mutation on miRNA
Background strain: 40D4
Plasmid transformation: MAR1 knock-down plasmid (hygromycin) transformed into hap2/FUS1R
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: HAP2-Flag in 40D
Background strain: 40D4
Plasmid transformation: HAP2-Flag-ble plasmid transformed into 40D4
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: Hsp-HAP2-HA in 63B10
Background strain: 63B10
Plasmid transformation: Hsp-HAP2-HA plasmid transformed into 63B10, co-transformation aphVIII
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: HAP2-CC662-3SS plasmid in 40D4, good expression level
Background strain: 40D4
Plasmid transformation: HAP2-CC662-3SS-aphVIII plasmid transformed into 40D4
Liu Y, Pei J, Grishin N, Snell WJ (2015) The cytoplasmic domain of the gamete membrane fusion protein HAP2 targets the protein to the fusion site in Chlamydomonas and regulates the fusion reaction. Development. 142:962-71
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: Cre12g541400 disrupted
Background strain: B215
Plasmid transformation: pSL72 plasmid transformed into B215 for mutagenesis
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: Cre12g541400 disrupted, generated by cross with 89G7 mt-
Background strain: B215
Plasmid transformation: pSL72 plasmid transformed into 21gr for mutagenesis, generated by cross with 89G7 mt-
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: HAP2-C661S in 40D4 (cell becomes defective on agglutination)
Background strain: 40D4
Plasmid transformation: HAP2-C661S plasmid transformed into 40D4, one plasmid
Liu Y, Pei J, Grishin N, Snell WJ (2015) The cytoplasmic domain of the gamete membrane fusion protein HAP2 targets the protein to the fusion site in Chlamydomonas and regulates the fusion reaction. Development. 142:962-71
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: hap2 mutant
Background strain: B215
Plasmid transformation: pSI103 plasmid transformed into B215, insertion mutagenesis on HAP2 gene
Liu Y, Tewari R, Ning J, Blagborough AM, Garbom S, Pei J, Grishin NV, Steele RE, Sinden RE, Snell WJ, Billker O (2008) The conserved plant sterility gene HAP2 functions after attachment of fusogenic membranes in Chlamydomonas and Plasmodium gametes. Genes Dev. 22:1051-68
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: HAP2-CC662-3SS plasmid in 40D4, mild expression level
Background strain: 40D4
Plasmid transformation: HAP2-CC662-3SS-aphVIII plasmid transformed into 40D4, one plasmid
Liu Y, Pei J, Grishin N, Snell WJ (2015) The cytoplasmic domain of the gamete membrane fusion protein HAP2 targets the protein to the fusion site in Chlamydomonas and regulates the fusion reaction. Development. 142:962-71
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: HAP2 N-terminal internal deletion in 40D4
Background strain: 40D4
Plasmid transformation: HAP2-NinteDele plasmid transformed into 40D4, one plasmid
Liu Y, Pei J, Grishin N, Snell WJ (2015) The cytoplasmic domain of the gamete membrane fusion protein HAP2 targets the protein to the fusion site in Chlamydomonas and regulates the fusion reaction. Development. 142:962-71
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: MAR1 (cre03g176961) insertion mutant at 3’UTR, from Chlamy library
Background strain: Jonikas CMJ030
Plasmid transformation: MAR1 (cre03g176961) insertion mutant at 3’UTR, from Chlamy library
CC-5319 Hap2/FUS1R-kd-20 mt-
$30.00
$30.00
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: MAR1-Flag knock-down cell
Background strain: 40D4
Plasmid transformation: MAR1 knock-down plasmid (hygromycin) transformed into hap2/FUS1R
CC-5320 Hap2/Fus1R-kd-15 mt-
$30.00
$30.00
From Yanjie Liu, William Snell lab, UT Southwestern Medical Center, May 2016
Mutant description: MAR1-Flag knock-down cell
Background strain: 40D4
Plasmid transformation: MAR1 knock-down plasmid (hygromycin) transformed into hap2/FUS1R
CC-5321 vtc1-1 mt-
$30.00
$30.00
From Sean Gallaher, Sabeeha Merchant group, UCLA, September 2016
Mutational insertion into strain CC-4425, followed by multiple crosses to CC-1690 and CC-1691. Sequence verified to have complete loss of vtc1 locus (Cre12.g510250), as well as the neighboring Cre12.g510252 locus.
Aksoy M, Pootakham W, and Grossman AR (2014) Critical Function of a Chlamydomonas reinhardtii Putative Polyphosphate Polymerase Subunit during Nutrient Deprivation. The Plant Cell. 26: 4214-4229
CC-5322 vtc4-1 mt-
$30.00
$30.00
From Sean Gallaher, Sabeeha Merchant group, UCLA, September 2016
Mutational insertion into strain CC-4425, followed by multiple crosses to CC-1690 and CC-1691. Sequence verified to have insertion into exon 1 of vtc4 locus (Cre09.g402775).
Aksoy M, Pootakham W, and Grossman AR (2014) Critical Function of a Chlamydomonas reinhardtii Putative Polyphosphate Polymerase Subunit during Nutrient Deprivation. The Plant Cell. 26: 4214-4229
From Sean Gallaher, Sabeeha Merchant group, UCLA, September 2016
vtc1-1 mutant strain (CC-5321) rescued with VTC1. Sequence verified to have insertion of VTC1 gene into the 3′ UTR of Cre14.g631250 locus.
Aksoy M, Pootakham W, and Grossman AR (2014) Critical Function of a Chlamydomonas reinhardtii Putative Polyphosphate Polymerase Subunit during Nutrient Deprivation. The Plant Cell. 26: 4214-4229
From Jacob Robertson, Jonikas lab, Princeton University, October 2016
This strain is a haploid progeny from a cross between 4A- (Dent, Haglund, Chin, Kobayashi, Niyogi, Plant Physiology 2005) and D66+ (Schnell and Lefebvre, Genetics 1993). It was selected from the progeny because it has a number of desirable properties: recovers well from cryogenic storage, electroporates well, mates well, grows well in the dark, grows well photoautotrophically, appears to have a functional carbon concentrating mechanism, does not clump much in liquid culture, swims normally and does not adhere much to glass.
This is the background strain used in the Chlamydomonas Library Project (www.chlamylibrary.org) and is the same as CC-4533 cw15 mt- [Jonikas CMJ030]. The only difference is that this isolate was thawed from cryogenic storage in October 2016.
Li X, Zhang R, Patena W, Gang SS, Blum SR, Ivanova N, Yue R, Robertson JM, Lefebvre PA, Fitz-Gibbon ST, Grossman AR, Jonikas MC (2016) An Indexed, Mapped Mutant Library Enables Reverse Genetics Studies of Biological Processes in Chlamydomonas reinhardtii. Plant Cell. 28:367-87
From Kelly Bower, Porter Lab, University of Minnesota, November 2016
This strain was created when pf2-4 mt- (CC-4483) was rescued with DRC4 with a D to K amino acid change at 198 carboxyl terminally tagged with GFP by transformation with pSI103 and selection on paromomycin.
Lewis WR, Malarkey EB, Tritschler D, Bower R, Pasek RC, Porath JD, Birket SE, Saunier S, Antignac C, Knowles MR, Leigh MW, Zariwala MA, Challa AK, Kesterson RA, Rowe SM, Drummond IA, Parant JM, Hildebrandt F, Porter E, Yoder BK, Berbari NF (2016) Mutation of Growth Arrest Specific 8 Reveals a Role in Motile Cilia Function and Human Disease. PLoS Genet. Jul 29;12(7):e1006220. doi:10.1371/journal.pgen.1006220. eCollection 2016 Jul. PubMed PMID: 27472056; PubMed Central PMCID: PMC4966937.
CC-5327 Osmo1 [Becker]
$30.00
$30.00
From Karin Komsic-Buchmann, Becker lab, University of Cologne, October 2016
Phenotype: CVs bigger
This strain cannot survive under osmotic conditions below 140 mosM.
Culture maintenance: photon flux ~20-70 mmol/m²s,14/10 hours light/dark cycle, 21°C, TAP + 0.2 g/l arginine + 64.6 mM sucrose
Komsic-Buchmann K, Stephan LM, Becker B (2012) The SEC6 protein is required for contractile vacuole function in Chlamydomonas reinhardtii. J Cell Sci 125:2885–2895
Komsic-Buchmann, Karin (2014) Structure and Function of the Contractile Vacuoles of Chlamdomonas reinhardtii. Dr. Hut, Munich, Germany.
CC-5328 Osmo2 [Becker]
$30.00
$30.00
From Karin Komsic-Buchmann, Becker lab, University of Cologne, October 2016
This strain cannot survive under osmotic conditions below 140 mosM.
Culture maintenance: photon flux ~20-70 mmol/m²s,14/10 hours light/dark cycle, 21°C, TAP + 0.2 g/l arginine + 64.6 mM sucrose
Komsic-Buchmann K, Stephan LM, Becker B (2012) The SEC6 protein is required for contractile vacuole function in Chlamydomonas reinhardtii. J Cell Sci 125:2885–2895
Komsic-Buchmann, Karin (2014) Structure and Function of the Contractile Vacuoles of Chlamdomonas reinhardtii. Dr. Hut, Munich, Germany.
CC-5329 Osmo4 [Becker]
$30.00
$30.00
From Karin Komsic-Buchmann, Becker lab, University of Cologne, October 2016
Phenotype: smaller CVs, shorter CV cycle, small cells
This strain cannot survive under osmotic conditions below 140 mosM.
Culture maintenance: photon flux ~20-70 mmol/m²s,14/10 hours light/dark cycle, 21°C, TAP + 0.2 g/l arginine + 64.6 mM sucrose
Komsic-Buchmann K, Stephan LM, Becker B (2012) The SEC6 protein is required for contractile vacuole function in Chlamydomonas reinhardtii. J Cell Sci 125:2885–2895
Komsic-Buchmann, Karin (2014) Structure and Function of the Contractile Vacuoles of Chlamdomonas reinhardtii. Dr. Hut, Munich, Germany.
CC-5330 Osmo6 [Becker]
$30.00
$30.00
From Karin Komsic-Buchmann, Becker lab, University of Cologne, October 2016
This strain cannot survive under osmotic conditions below 140 mosM.
Culture maintenance: photon flux ~20-70 mmol/m²s,14/10 hours light/dark cycle, 21°C, TAP + 0.2 g/l arginine + 64.6 mM sucrose
Komsic-Buchmann K, Stephan LM, Becker B (2012) The SEC6 protein is required for contractile vacuole function in Chlamydomonas reinhardtii. J Cell Sci 125:2885–2895
Komsic-Buchmann, Karin (2014) Structure and Function of the Contractile Vacuoles of Chlamdomonas reinhardtii. Dr. Hut, Munich, Germany.
CC-5331 Osmo7 [Becker]
$30.00
$30.00
From Karin Komsic-Buchmann, Becker lab, University of Cologne, October 2016
Phenotype: longer systole phase
This strain cannot survive under osmotic conditions below 140 mosM.
Culture maintenance: photon flux ~20-70 mmol/m²s,14/10 hours light/dark cycle, 21°C, TAP + 0.2 g/l arginine + 64.6 mM sucrose
Komsic-Buchmann K, Stephan LM, Becker B (2012) The SEC6 protein is required for contractile vacuole function in Chlamydomonas reinhardtii. J Cell Sci 125:2885–2895
Komsic-Buchmann, Karin (2014) Structure and Function of the Contractile Vacuoles of Chlamdomonas reinhardtii. Dr. Hut, Munich, Germany.
CC-5332 Osmo9 [Becker]
$30.00
$30.00
From Karin Komsic-Buchmann, Becker lab, University of Cologne, October 2016
Phenotype: CVs bigger, higher efflux
This strain cannot survive under osmotic conditions below 140 mosM.
Culture maintenance: photon flux ~20-70 mmol/m²s,14/10 hours light/dark cycle, 21°C, TAP + 0.2 g/l arginine + 64.6 mM sucrose
Komsic-Buchmann K, Stephan LM, Becker B (2012) The SEC6 protein is required for contractile vacuole function in Chlamydomonas reinhardtii. J Cell Sci 125:2885–2895
Komsic-Buchmann, Karin (2014) Structure and Function of the Contractile Vacuoles of Chlamdomonas reinhardtii. Dr. Hut, Munich, Germany.
CC-5333 Osmo10 [Becker]
$30.00
$30.00
From Karin Komsic-Buchmann, Becker lab, University of Cologne, October 2016
Phenotype: CVs bigger, higher efflux
This strain cannot survive under osmotic conditions below 140 mosM.
Culture maintenance: photon flux ~20-70 mmol/m²s,14/10 hours light/dark cycle, 21°C, TAP + 0.2 g/l arginine + 64.6 mM sucrose
Komsic-Buchmann K, Stephan LM, Becker B (2012) The SEC6 protein is required for contractile vacuole function in Chlamydomonas reinhardtii. J Cell Sci 125:2885–2895
Komsic-Buchmann, Karin (2014) Structure and Function of the Contractile Vacuoles of Chlamdomonas reinhardtii. Dr. Hut, Munich, Germany.
CC-5334 Osmo11 [Becker]
$30.00
$30.00
From Karin Komsic-Buchmann, Becker lab, University of Cologne, October 2016
Phenotype: smaller CVs, normal CV cycle
This strain cannot survive under osmotic conditions below 140 mosM.
Culture maintenance: photon flux ~20-70 mmol/m²s,14/10 hours light/dark cycle, 21°C, TAP + 0.2 g/l arginine + 64.6 mM sucrose
Komsic-Buchmann K, Stephan LM, Becker B (2012) The SEC6 protein is required for contractile vacuole function in Chlamydomonas reinhardtii. J Cell Sci 125:2885–2895
Komsic-Buchmann, Karin (2014) Structure and Function of the Contractile Vacuoles of Chlamdomonas reinhardtii. Dr. Hut, Munich, Germany.
CC-5335 Osmo18 [Becker]
$30.00
$30.00
From Karin Komsic-Buchmann, Becker lab, University of Cologne, October 2016
Phenotype: CVs smaller, CV cycle shorter, less efflux
This strain cannot survive under osmotic conditions below 140 mosM.
Culture maintenance: photon flux ~20-70 mmol/m²s,14/10 hours light/dark cycle, 21°C, TAP + 0.2 g/l arginine + 64.6 mM sucrose
Komsic-Buchmann K, Stephan LM, Becker B (2012) The SEC6 protein is required for contractile vacuole function in Chlamydomonas reinhardtii. J Cell Sci 125:2885–2895
Komsic-Buchmann, Karin (2014) Structure and Function of the Contractile Vacuoles of Chlamdomonas reinhardtii. Dr. Hut, Munich, Germany.
CC-5336 Osmo20 [Becker]
$30.00
$30.00
From Karin Komsic-Buchmann, Becker lab, University of Cologne, October 2016
Phenotype: smaller CVs
This strain cannot survive under osmotic conditions below 140 mosM.
Culture maintenance: photon flux ~20-70 mmol/m²s,14/10 hours light/dark cycle, 21°C, TAP + 0.2 g/l arginine + 64.6 mM sucrose
Komsic-Buchmann K, Stephan LM, Becker B (2012) The SEC6 protein is required for contractile vacuole function in Chlamydomonas reinhardtii. J Cell Sci 125:2885–2895
Komsic-Buchmann, Karin (2014) Structure and Function of the Contractile Vacuoles of Chlamdomonas reinhardtii. Dr. Hut, Munich, Germany.
CC-5337 Osmo21 [Becker]
$30.00
$30.00
From Karin Komsic-Buchmann, Becker lab, University of Cologne, October 2016
Phenotype: CV cycle variable
This strain cannot survive under osmotic conditions below 140 mosM.
Culture maintenance: photon flux ~20-70 mmol/m²s,14/10 hours light/dark cycle, 21°C, TAP + 0.2 g/l arginine + 64.6 mM sucrose
Komsic-Buchmann K, Stephan LM, Becker B (2012) The SEC6 protein is required for contractile vacuole function in Chlamydomonas reinhardtii. J Cell Sci 125:2885–2895
Komsic-Buchmann, Karin (2014) Structure and Function of the Contractile Vacuoles of Chlamdomonas reinhardtii. Dr. Hut, Munich, Germany.
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