Articoli di riviste: "Chlamydomonas reinhardtii – Genetics"

1

Roesler, Keith R., e William L. Ogren. "Chlamydomonas reinhardtii Phosphoribulokinase". Plant Physiology 93, n. 1 (1 maggio 1990): 188–93. http://dx.doi.org/10.1104/pp.93.1.188.

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2

Stauber, Einar J., e Michael Hippler. "Chlamydomonas reinhardtii proteomics". Plant Physiology and Biochemistry 42, n. 12 (dicembre 2004): 989–1001. http://dx.doi.org/10.1016/j.plaphy.2004.09.008.

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3

Lamb, Mary Rose, Susan K. Dutcher, Cathy K. Worley e Carol L. Dieckmann. "Eyespot-Assembly Mutants in Chlamydomonas reinhardtii". Genetics 153, n. 2 (1 ottobre 1999): 721–29. http://dx.doi.org/10.1093/genetics/153.2.721.

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Abstract Chlamydomonas reinhardtii is a single-celled green alga that phototaxes toward light by means of a light-sensitive organelle, the eyespot. The eyespot is composed of photoreceptor and Ca++-channel signal transduction components in the plasma membrane of the cell and reflective carotenoid pigment layers in an underlying region of the large chloroplast. To identify components important for the positioning and assembly of a functional eyespot, a large collection of nonphototactic mutants was screened for those with aberrant pigment spots. Four loci were identified. eye2 and eye3 mutants have no pigmented eyespots. min1 mutants have smaller than wild-type eyespots. mlt1(ptx4) mutants have multiple eyespots. The MIN1, MLT1(PTX4), and EYE2 loci are closely linked to each other; EYE3 is unlinked to the other three loci. The eye2 and eye3 mutants are epistatic to min1 and mlt1 mutations; all double mutants are eyeless. min1 mlt1 double mutants have a synthetic phenotype; they are eyeless or have very small, misplaced eyespots. Ultrastructural studies revealed that the min1 mutants are defective in the physical connection between the plasma membrane and the chloroplast envelope membranes in the region of the pigment granules. Characterization of these four loci will provide a beginning for the understanding of eyespot assembly and localization in the cell.

4

Kuchka, Michael R., e Jonathan W. Jarvik. "Short-Flagella Mutants of Chlamydomonas reinhardtii". Genetics 115, n. 4 (1 aprile 1987): 685–91. http://dx.doi.org/10.1093/genetics/115.4.685.

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ABSTRACT Six short-flagella mutants were isolated by screening clones of mutagenized Chlamydomonas for slow swimmers. The six mutants identify three unlinked Mendelian genes, with three mutations in gene shf-1, two in shf-2 and one in shf-3. shf-1 and shf-2 have been mapped to chromosomes VI and I, respectively. Two of the shf-1 mutations have temperature-sensitive flagellar-assembly phenotypes, and one shf-2 mutant has a cold-sensitive phenotype. shf shf double mutants were constructed; depending on the alleles present they showed either flagellaless or short-flagella phenotypes. Phenotypic revertants of shf-1 and shf-2 mutants were isolated, and certain of them were found to carry extragenic suppressors, some dominant and some recessive. We suspect that the shf mutations affect components of a specific flagellar size-control system, the existence of which has been suggested by a variety of physiological experiments.

5

Porter, Mary E., Julie A. Knott, Steven H. Myster e Samuel J. Farlow. "The Dynein Gene Family in Chlamydomonas reinhardtii". Genetics 144, n. 2 (1 ottobre 1996): 569–85. http://dx.doi.org/10.1093/genetics/144.2.569.

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Abstract To correlate dynein heavy chain (Dhc) genes with flagellar mutations and gain insight into the function of specific dynein isoforms, we placed eight members of the Dhc gene family on the genetic map of Chlamydomonas. Using a PCR-based strategy, we cloned 11 Dhc genes from Chlamydomonas. Comparisons with other Dhc genes indicate that two clones correspond to genes encoding the alpha and beta heavy chains of the outer dynein arm. Alignment of the predicted amino acid sequences spanning the nucleotide binding site indicates that the remaining nine clones can be subdivided into three groups that are likely to include representatives of the inner-arm Dhc isoforms. Gene-specific probes reveal that each clone represents a single-copy gene that is expressed as a transcript of the appropriate size (>13 kb) sufficient to encode a high molecular weight Dhc polypeptide. The expression of all nine genes is upregulated in response to deflagellation, suggesting a role in axoneme assembly or motility. Restriction fragment length polymorphisms between divergent C. reinhardtii strains have been used to place each Dhr gene on the genetic map of Chlamydomonas. These studies lay the groundwork for correlating defects in different Dhc genes with specific flagellar mutations.

6

Shimogawara, Kosuke, Shoko Fujiwara, Arthur Grossman e Hideaki Usuda. "High-Efficiency Transformation of Chlamydomonas reinhardtii by Electroporation". Genetics 148, n. 4 (1 aprile 1998): 1821–28. http://dx.doi.org/10.1093/genetics/148.4.1821.

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Abstract We have established a high-efficiency method for transforming the unicellular, green alga Chlamydomonas reinhardtii by electroporation. Electroporation of strains CC3395 and CC425, cell wall-less mutants devoid of argininosuccinate lyase (encoded by ARG7), in the presence of the plasmid pJD67 (which contains ARG7) was used to optimize conditions for the introduction of exogenous DNA. The conditions that were varied included osmolarity, temperature, concentration of exogenous DNA, voltage and capacitance. Following optimization, the maximum transformation frequency obtained was 2 × 105 transformants per μg of DNA; this frequency is two orders of magnitude higher than obtained with the current standard method using glass beads to introduce exogenous DNA. The electroporation procedure described in this article is of general utility, and makes it feasible to isolate genes by direct complementation of Chlamydomonas reinhardtii mutants.

7

Dutcher, S. K., R. E. Galloway, W. R. Barclay e G. Poortinga. "Tryptophan analog resistance mutations in Chlamydomonas reinhardtii." Genetics 131, n. 3 (1 luglio 1992): 593–607. http://dx.doi.org/10.1093/genetics/131.3.593.

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Abstract Forty single gene mutations in Chlamydomonas reinhardtii were isolated based on resistance to the compound 5'-methyl anthranilic acid (5-MAA). In other organisms, 5-MAA is converted to 5'-methyltryptophan (5-MT) and 5-MT is a potent inhibitor of anthranilate synthase, which catalyzes the first committed step in tryptophan biosynthesis. The mutant strains fall into two phenotypic classes based on the rate of cell division in the absence of 5-MAA. Strains with class I mutations divide more slowly than wild-type cells. These 17 mutations map to seven loci, which are designated MAA1 to MAA7. Strains with class II mutations have generation times indistinguishable from wild-type cells, and 7 of these 23 mutations map to loci defined by class I mutations. The remainder of the class II mutations map to 9 other loci, which are designated MAA8-MAA16. The maa5-1 mutant strain excretes high levels of anthranilate and phenylalanine into the medium. In this strain, four enzymatic activities in the tryptophan biosynthetic pathway are increased at least twofold. These include the combined activities of anthranilate phosphoribosyl transferase, phosphoribosyl anthranilate isomerase, indoleglycerol phosphate synthetase and anthranilate synthase. The slow growth phenotypes of strains with class I mutations are not rescued by the addition of tryptophan, but the slow growth phenotype of the maa6-1 mutant strain is partially rescued by the addition of indole. The maa6-1 mutant strain excretes a fluorescent compound into the medium, and cell extracts have no combined anthranilate phosphoribosyl transferase, phosphoribosyl anthranilate isomerase and indoleglycerol phosphate synthetase activity. The MAA6 locus is likely to encode a tryptophan biosynthetic enzyme. None of the other class I mutations affected these enzyme activities. Based on the phenotypes of double mutant strains, epistatic relationships among the class I mutations have been determined.

8

Grossman, Arthur R. "Chlamydomonas reinhardtii and photosynthesis: genetics to genomics". Current Opinion in Plant Biology 3, n. 2 (aprile 2000): 132–37. http://dx.doi.org/10.1016/s1369-5266(99)00053-9.

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9

Ferris, P. J. "Characterization of a Chlamydomonas transposon, Gulliver, resembling those in higher plants." Genetics 122, n. 2 (1 giugno 1989): 363–77. http://dx.doi.org/10.1093/genetics/122.2.363.

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Abstract While pursuing a chromosomal walk through the mt+ locus of linkage group VI of Chlamydomonas reinhardtii, I encountered a 12-kb sequence that was found to be present in approximately 12 copies in the nuclear genome. Comparison of various C. reinhardtii laboratory strains provided evidence that the sequence was mobile and therefore a transposon. One of two separate natural isolates interfertile with C. reinhardtii, C. smithii (CC-1373), contained the transposon, but at completely different locations in its nuclear genome than C. reinhardtii; and a second, CC-1952 (S1-C5), lacked the transposon altogether. Genetic analysis indicated that the transposon was found at dispersed sites throughout the genome, but had a conserved structure at each location. Sequence homology between the termini was limited to an imperfect 15-bp inverted repeat. An 8-bp target site duplication was created by insertion; transposon sequences were completely removed upon excision leaving behind both copies of the target site duplication, with minor base changes. The transposon contained an internal region of unique repetitive sequence responsible for restriction fragment length heterogeneity among the various copies of the transposon. In several cases it was possible to identify which of the dozen transposons in a given strain served as the donor when a transposition event occurred. The transposon often moved into a site genetically linked to the donor, and transposition appeared to be nonreplicative. Thus the mechanism of transposition and excision of the transposon, which I have named Gulliver, resembles that of certain higher plant transposons, like the Ac transposon of maize.

10

Bennoun, P., M. Delosme e U. Kück. "Mitochondrial genetics of Chlamydomonas reinhardtii: resistance mutations marking the cytochrome b gene." Genetics 127, n. 2 (1 febbraio 1991): 335–43. http://dx.doi.org/10.1093/genetics/127.2.335.

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Abstract We describe the genetic and molecular analysis of the first non-Mendelian mutants of Chlamydomonas reinhardtii resistant to myxothiazol, an inhibitor of the respiratory cytochrome bc1 complex. Using a set of seven oligonucleotide probes, restriction fragments containing the mitochondrial cytochrome b (cyt b) gene from C. reinhardtii were isolated from a mitochondrial DNA library. This gene is located adjacent to the gene for subunit 4 of the mitochondrial NADH-dehydrogenase (ND4), near one end of the 15.8-kb linear mitochondrial genome of C. reinhardtii. The algal cytochrome b apoprotein contains 381 amino-acid residues and exhibits a sequence similarity of about 59% with other plant cytochrome b proteins. The cyt b gene from four myxothiazol resistant mutants of C. reinhardtii was amplified for DNA sequence analysis. In comparison to the wild-type strain, all mutants contain an identical point mutation in the cyt b gene, leading to a change of a phenylalanine codon to a leucine codon at amino acid position 129 of the cytochrome b protein. Segregation analysis in tetrads from reciprocal crosses of mutants with wild type shows a strict uniparental inheritance of this mutation from the mating type minus parent (UP-). However, mitochondrial markers from both parents are recovered in vegetative diploids in variable proportions from one experiment to the next for a given cross. On the average, a strong bias is seen for markers inherited from the mating type minus parent.

11

Beck, Christoph F., e Axel Acker. "Gametic Differentiation of Chlamydomonas reinhardtii". Plant Physiology 98, n. 3 (1 marzo 1992): 822–26. http://dx.doi.org/10.1104/pp.98.3.822.

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12

Tam, L. W., e P. A. Lefebvre. "Cloning of flagellar genes in Chlamydomonas reinhardtii by DNA insertional mutagenesis." Genetics 135, n. 2 (1 ottobre 1993): 375–84. http://dx.doi.org/10.1093/genetics/135.2.375.

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Abstract Chlamydomonas is a popular genetic model system for studying many cellular processes. In this report, we describe a new approach to isolate Chlamydomonas genes using the cloned nitrate reductase gene (NIT1) as an insertional mutagen. A linearized plasmid containing the NIT1 gene was introduced into nit1 mutant cells by glass-bead transformation. Of 3000 Nit+ transformants examined, 74 showed motility defects of a wide range of phenotypes, suggesting that DNA transformation is an effective method for mutagenizing cells. For 13 of 15 such motility mutants backcrossed to nit- mutant strains, the motility phenotype cosegregated with the Nit+ phenotype, indicating that the motility defects of these 13 mutants may be caused by integration of the plasmid. Further genetic analysis indicated that three of these mutants contained alleles of previously identified loci: mbo2 (move backward only), pf13 (paralyzed flagella) and vfl1 (variable flagellar number). Three other abnormal-flagellar-number mutants did not map to any previously described loci at which mutations produce similar phenotypes. Genomic sequences flanking the integrated plasmid in the mbo2 and vfl1 mutants were isolated and used as probes to obtain wild-type genomic clones, which complemented the motility defects upon transformation into cells. Our results demonstrate the potential of this new approach for cloning genes identified by mutation in Chlamydomonas.

13

Herron, Matthew D., William C. Ratcliff, Jacob Boswell e Frank Rosenzweig. "Genetics of a de novo origin of undifferentiated multicellularity". Royal Society Open Science 5, n. 8 (agosto 2018): 180912. http://dx.doi.org/10.1098/rsos.180912.

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The evolution of multicellularity was a major transition in evolution and set the stage for unprecedented increases in complexity, especially in land plants and animals. Here, we explore the genetics underlying a de novo origin of multicellularity in a microbial evolution experiment carried out on the green alga Chlamydomonas reinhardtii . We show that large-scale changes in gene expression underlie the transition to a multicellular life cycle. Among these, changes to genes involved in cell cycle and reproductive processes were overrepresented, as were changes to C. reinhardtii -specific and volvocine-specific genes. These results suggest that the genetic basis for the experimental evolution of multicellularity in C. reinhardtii has both lineage-specific and shared features, and that the shared features have more in common with C. reinhardtii 's relatives among the volvocine algae than with other multicellular green algae or land plants.

14

Dayer, Régine, Beat B. Fischer, Rik I. L. Eggen e Stéphane D. Lemaire. "The Peroxiredoxin and Glutathione Peroxidase Families in Chlamydomonas reinhardtii". Genetics 179, n. 1 (maggio 2008): 41–57. http://dx.doi.org/10.1534/genetics.107.086041.

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15

Figueroa-Martínez, Francisco, Soledad Funes, Lars-Gunnar Franzén e Diego González-Halphen. "Reconstructing the Mitochondrial Protein Import Machinery of Chlamydomonas reinhardtii". Genetics 179, n. 1 (maggio 2008): 149–55. http://dx.doi.org/10.1534/genetics.108.087965.

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16

Ness, Rob W., Andrew D. Morgan, Nick Colegrave e Peter D. Keightley. "Estimate of the Spontaneous Mutation Rate in Chlamydomonas reinhardtii". Genetics 192, n. 4 (10 ottobre 2012): 1447–54. http://dx.doi.org/10.1534/genetics.112.145078.

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17

Barsel, S. E., D. E. Wexler e P. A. Lefebvre. "Genetic analysis of long-flagella mutants of Chlamydomonas reinhardtii." Genetics 118, n. 4 (1 aprile 1988): 637–48. http://dx.doi.org/10.1093/genetics/118.4.637.

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Abstract The length of the flagella of Chlamydomonas reinhardtii cells is tightly regulated; both short-flagella and long-flagella mutants have been described. This report characterizes ten long-flagella mutants, including five newly isolated mutants, to determine the number of different loci conferring this phenotype, and to study interactions of mutants at different loci. The mutants, each of which was recessive in heterozygous diploids with wild type, fall into three unlinked complementation groups. One of these defines a new gene, lf3, which maps near the centromere of linkage group I. The flagellar length distributions in populations of each mutant were broad, with the longest flagella measuring four times the length of the longest flagella seen on wild-type cells. Each of the ten mutants had defective flagellar regrowth after amputation. Some of the mutants showed no regrowth within the time required for wild-type cells to regenerate flagella completely. Other mutants had subpopulations with rapid regeneration kinetics, and subpopulations with no observable regeneration. The mutants were each crossed to wild type to form temporary quadriflagellate, dikaryon cells; in each case the long flagella were rapidly shortened in the presence of the wild-type cytoplasm, demonstrating that the mutants were recessive, and that length control could be exerted on already assembled flagella.

18

Podstavková, S., E. Miadoková e D. Vlček. "Repair-deficient mutants of Chlamydomonas reinhardtii". Mutation Research/Environmental Mutagenesis and Related Subjects 271, n. 2 (1992): 150. http://dx.doi.org/10.1016/0165-1161(92)91168-q.

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19

Remacle, C., F. Duby, P. Cardol e R. F. Matagne. "Mutations inactivating mitochondrial genes in Chlamydomonas reinhardtii". Biochemical Society Transactions 29, n. 4 (1 agosto 2001): 442–46. http://dx.doi.org/10.1042/bst0290442.

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Chlamydomonas reinhardtii is now becoming a useful model for the study of mitochondrial genetics in a photosynthetic organism. The small (15.8 kb) mitochondrial genome C. reinhardtii has been sequenced completely and all the genes have been identified. Several mutants inactivated in mitochondrial genes encoding components of the respiratory complexes I, III and IV have been characterized at the molecular level. Assembly of complex I in several mutant strains and mapping of mitochondrial mutations by recombinational analysis are also described.

20

Goldschmidt-Clermont, Michel, Jacqueline Girard-Bascou, Yves Choquet e Jean-David Rochaix. "Trans-splicing mutants of Chlamydomonas reinhardtii". Molecular and General Genetics MGG 223, n. 3 (settembre 1990): 417–25. http://dx.doi.org/10.1007/bf00264448.

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21

Mukherjee, Ananya. "State Transition Regulation in Chlamydomonas reinhardtii". Plant Physiology 183, n. 4 (agosto 2020): 1418–19. http://dx.doi.org/10.1104/pp.20.00814.

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22

Suzuki, Kensaku, Laura Fredrick Marek e Martin H. Spalding. "A Photorespiratory Mutant of Chlamydomonas reinhardtii". Plant Physiology 93, n. 1 (1 maggio 1990): 231–37. http://dx.doi.org/10.1104/pp.93.1.231.

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23

Klein, Uwe. "Intracellular Carbon Partitioning in Chlamydomonas reinhardtii". Plant Physiology 85, n. 4 (1 dicembre 1987): 892–97. http://dx.doi.org/10.1104/pp.85.4.892.

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24

Manuel, Livingston J., e James V. Moroney. "Inorganic Carbon Accumulation by Chlamydomonas reinhardtii". Plant Physiology 88, n. 2 (1 ottobre 1988): 491–96. http://dx.doi.org/10.1104/pp.88.2.491.

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25

Muñoz-Blanco, J. "Glutamate dehydrogenase isozymes of Chlamydomonas reinhardtii". FEMS Microbiology Letters 61, n. 3 (15 ottobre 1989): 315–18. http://dx.doi.org/10.1016/0378-1097(89)90217-6.

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26

Holmes, J. A., D. E. Johnson e S. K. Dutcher. "Linkage group XIX of Chlamydomonas reinhardtii has a linear map." Genetics 133, n. 4 (1 aprile 1993): 865–74. http://dx.doi.org/10.1093/genetics/133.4.865.

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Abstract Linkage group XIX (or the UNI linkage group) of Chlamydomonas reinhardtii has been reported to show a circular meiotic recombination map. A circular map predicts the existence of strong chiasma and chromatid interference, which would lead to an excess number of two-strand double crossovers during meiosis. We have tested this prediction in multipoint crosses. Our results are consistent with a linear linkage group that shows positive chiasma interference and no chromatid interference. Chiasma interference occurs both within arms and across the centromere. Of the original loci that contributed to the circular map, we find that two map to other linkage groups and a third cannot be retested because the mutant strain that defined it has been lost. A second reported unusual property for linkage group XIX was the increase in meiotic recombination with increases in temperature during a period that precedes the onset of meiosis. Although we observed changes in recombination frequencies in some intervals on linkage group XIX in crosses to CC-1952, and in strains heterozygous for the mutation ger1 at 16 degrees, we also show that our strains do not exhibit the previously observed patterns of temperature-sensitive recombination for two different pairs of loci on linkage group XIX. We conclude that linkage group XIX has a linear genetic map that is not significantly different from other Chlamydomonas linkage groups.

27

McCarthy, Sarah S., Marilyn C. Kobayashi e Krishna K. Niyogi. "White Mutants of Chlamydomonas reinhardtii Are Defective in Phytoene Synthase". Genetics 168, n. 3 (novembre 2004): 1249–57. http://dx.doi.org/10.1534/genetics.104.030635.

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Ferris, Patrick J., E. Virginia Armbrust e Ursula W. Goodenough. "Genetic Structure of the Mating-Type Locus of Chlamydomonas reinhardtii". Genetics 160, n. 1 (1 gennaio 2002): 181–200. http://dx.doi.org/10.1093/genetics/160.1.181.

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Abstract Portions of the cloned mating-type (MT) loci (mt+ and mt−) of Chlamydomonas reinhardtii, defined as the ~1-Mb domains of linkage group VI that are under recombinational suppression, were subjected to Northern analysis to elucidate their coding capacity. The four central rearranged segments of the loci were found to contain both housekeeping genes (expressed during several life-cycle stages) and mating-related genes, while the sequences unique to mt+ or mt− carried genes expressed only in the gametic or zygotic phases of the life cycle. One of these genes, Mtd1, is a candidate participant in gametic cell fusion; two others, Mta1 and Ezy2, are candidate participants in the uniparental inheritance of chloroplast DNA. The identified housekeeping genes include Pdk, encoding pyruvate dehydrogenase kinase, and GdcH, encoding glycine decarboxylase complex subunit H. Unusual genetic configurations include three genes whose sequences overlap, one gene that has inserted into the coding region of another, several genes that have been inactivated by rearrangements in the region, and genes that have undergone tandem duplication. This report extends our original conclusion that the MT locus has incurred high levels of mutational change.

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Hodson, R. C., e P. M. Gresshoff. "Fluoroacetamide resistance mutations in Chlamydomonas reinhardtii". Archives of Microbiology 148, n. 1 (giugno 1987): 8–13. http://dx.doi.org/10.1007/bf00429639.

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30

Gloeckner, G., e C. F. Beck. "Genes involved in light control of sexual differentiation in Chlamydomonas reinhardtii." Genetics 141, n. 3 (1 novembre 1995): 937–43. http://dx.doi.org/10.1093/genetics/141.3.937.

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Abstract Gamete formation requires the sequential action of two extrinsic cues, nitrogen deprivation and blue light. The mutants described here are specifically altered in the light-dependent step. Mutations lrg1, lrg3, and lrg4 overcome this light dependence while mutation lrg2 results in a delayed execution of the light-mediated step. The four mutations are linked. The recessive nature of the lrg1, lrg3, and lrg4 mutations implies that they encode elements of negative control in this light response pathway. Analyses of diploids suggest an interaction between the gene products of the mutated loci with a central role for lrg4. The lrg4 mutation is unique also because it overcomes the light dependence of Chlamydomonas zygote germination when present in homozygous form. These data indicate that there are common components in the signal chains that control gametogenesis and zygote germination.

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Moroney, James V., Barbara J. Wilson e N. E. Tolbert. "Glycolate Metabolism and Excretion by Chlamydomonas reinhardtii". Plant Physiology 82, n. 3 (1 novembre 1986): 821–26. http://dx.doi.org/10.1104/pp.82.3.821.

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Newman, S. M., E. H. Harris, A. M. Johnson, J. E. Boynton e N. W. Gillham. "Nonrandom distribution of chloroplast recombination events in Chlamydomonas reinhardtii: evidence for a hotspot and an adjacent cold region." Genetics 132, n. 2 (1 ottobre 1992): 413–29. http://dx.doi.org/10.1093/genetics/132.2.413.

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Abstract Intermolecular recombination of Chlamydomonas chloroplast genes has been analyzed in sexual crosses and following biolistic transformation. The pattern and position of specific exchange events within 15 kb of the 22-kb inverted repeat have been mapped with respect to known restriction fragment length polymorphism markers that distinguish the chloroplast genomes of the interfertile species Chlamydomonas reinhardtii and Chlamydomonas smithii. Recombinant progeny were selected from two- and three-factor crosses involving point mutations conferring herbicide (dr) and antibiotic resistance (er and spr) in the psbA, 23S and 16S ribosomal RNA genes, respectively. Exchange events were not randomly distributed over the 15-kb region, but were found to occur preferentially in a 0.7-kb sequence spanning the 3' end of the psbA gene and were much less common in an adjacent region of ca. 2.0 kb. These findings are corroborated by data showing that the dr mutation is unlinked genetically (3% recombination/kb) to the er and spr rRNA mutations, which are themselves linked and show ca. 1% recombination/kb. This discrepancy is significant since the dr-er and er-spr intervals are about the same length (ca. 7 kb). During chloroplast transformation, the 0.7-kb recombination hotspot also functions as a preferential site for exchange events leading to the integration of donor psbA gene sequences. The 0.7-kb hotspot region contains four classes of 18-37-bp direct repeats also found in other intergenic regions, but no open reading frame. Using deletion constructs in a chloroplast transformation assay, the hotspot was localized to a 500-bp region that lacks most of these repeats, which suggests that the repeats themselves are not responsible for the increased recombination frequency. Within this region, a 400-bp sequence is highly conserved between the chloroplast genomes of C. reinhardtii and C. smithii and includes several structural motifs characteristic of recombination hotspots in other systems.

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Virolainen, Pavel A., e Elena M. Chekunova. "Optimization of CRISPR/Cas9 method for transgenesis of model microalgae <i>Chlamydomonas reinhardtii</i>". Ecological genetics 20, n. 1S (8 dicembre 2022): 42–43. http://dx.doi.org/10.17816/ecogen112332.

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In this work we knocked out the LTS3 gene of the microalgae Chlamydomonas reinhardtii using the TIM technique optimized for the available equipment. We achieved transformation efficiency of 68.8%, knockout of this gene lead to the death of C. reinhardtii cells after several division cycles. The creation and study of genetically modified organisms in fundamental research allows a deeper understanding of the basic processes in the cells with the prospect of further applying this knowledge in practice. Microalgae are an interesting object for genetic engineering because of the great prospects for their application in biotechnology, but in almost every case it is necessary to develop new strategies and transformation methods for the introduction of genetic constructs into the cell. CRISPR/Cas revolutionized the field of genome editing due to its simplicity, efficiency and accuracy compared to previously used methods, which over time simplified the development of protocols [1]. Currently, the most effective method of transformation is TIM (Targeted Insertional Mutagenesis) [2], developed for the microalgae Chlamydomonas reinhardtii P.A. Dang. model object of photosynthesis genetics. To test and optimize the TIM technique [2] in our lab, we carried out a knockout of the LTS3 gene, a transcriptional activator of chlorophyll biosynthesis genes in heterotrophic conditions [3]. We used glass beads agitation and electroporation (Gene Pulser Xcell, Bio-Rad, USA) methods in order to introduce into C. reinhardtii cells of the CC-125 (wt, mt+) strain the ribonucleoprotein complex SpCas9/sgRNA and double-stranded donor DNA with paromomycin resistance gene. The effectiveness of transformation varied from 10.6% to 68.8%. Probably, the LTS3 gene product plays a key role in the pathway of chlorophyll biosynthesis, since its knockout led to the death of C. reinhardtii cells after several division cycles. The transformation protocol optimized for the equipment available in our lab can be further refined and used to study the functions of other C. reinhardtii genes.

34

Lee, Robert W., e Claude Lemieux. "BIPARENTAL INHERITANCE OF NON-MENDELIAN GENE MARKERS IN CHLAMYDOMONAS MOEWUSII". Genetics 113, n. 3 (1 luglio 1986): 589–600. http://dx.doi.org/10.1093/genetics/113.3.589.

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ABSTRACT The first two non-Mendelian gene mutations to be identified in Chlamydomonas moewusii are described. These putative chloroplast gene mutations include one for resistance to streptomycin (sr-nM1) and one for resistance to erythromycin (er-nM1). In one- and two-factor reciprocal crosses, usually over 90% of the germinating zygospores transmitted these mutations and their wild-type alternatives from both parents (biparental zygospores); the remaining zygospores transmitted exclusively the non-Mendelian markers of the mating-type "plus" parent. Among the biparental zygospores, a strong bias in the transmission of non-Mendelian alleles from the mating-type "plus" parent was indicated by an excess of meiotic and postmeiotic mitotic progeny that were homoplasmic for non-Mendelian alleles from this parent compared to those that were homoplasmic for the non-Mendelian alleles from the mating-type "minus" parent. At best, weak linkage was detected between the sr-nM1 and er-nM1 loci. Non-Mendelian, chloroplast gene markers in Chlamydomonas eugametos and Chlamydomonas reinhardtii showed a predominantly uniparental mode of transmission from the mating-type "plus" parent in crosses performed under the same conditions used for the C. moewusii crosses.

35

Mahan, Kristina M., Obed W. Odorn e David L. Herrin. "Controlling fungal contamination in Chlamydomonas reinhardtii cultures". BioTechniques 39, n. 4 (ottobre 2005): 457–58. http://dx.doi.org/10.2144/000112022.

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36

Schimmer, Oskar, e Irmgard Kühne. "Furoquinoline alkaloids as photosensitizers in Chlamydomonas reinhardtii". Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 249, n. 1 (luglio 1991): 105–10. http://dx.doi.org/10.1016/0027-5107(91)90136-c.

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37

Loppes, Roland, e Ralph Heindricks. "New arginine-requiring mutants in Chlamydomonas reinhardtii". Archives of Microbiology 143, n. 4 (gennaio 1986): 348–52. http://dx.doi.org/10.1007/bf00412801.

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38

Roberts, Douglas G. W., Mary Rose Lamb e Carol L. Dieckmann. "Characterization of the EYE2 Gene Required for Eyespot Assembly in Chlamydomonas reinhardtii". Genetics 158, n. 3 (1 luglio 2001): 1037–49. http://dx.doi.org/10.1093/genetics/158.3.1037.

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Abstract The unicellular biflagellate green alga Chlamydomonas reinhardtii can perceive light and respond by altering its swimming behavior. The eyespot is a specialized structure for sensing light, which is assembled de novo at every cell division from components located in two different cellular compartments. Photoreceptors and associated signal transduction components are localized in a discrete patch of the plasma membrane. This patch is tightly packed against an underlying sandwich of chloroplast membranes and carotenoid-filled lipid granules, which aids the cell in distinguishing light direction. In a prior screen for mutant strains with eyespot defects, the EYE2 locus was defined by the single eye2-1 allele. The mutant strain has no eyespot by light microscopy and has no organized carotenoid granule layers as judged by electron microscopy. Here we demonstrate that the eye2-1 mutant is capable of responding to light, although the strain is far less sensitive than wild type to low light intensities and orients imprecisely. Therefore, pigment granule layer assembly in the chloroplast is not required for photoreceptor localization in the plasma membrane. A plasmid-insertion mutagenesis screen yielded the eye2-2 allele, which allowed the isolation and characterization of the EYE2 gene. The EYE2 protein is a member of the thioredoxin superfamily. Site-directed mutagenesis of the active site cysteines demonstrated that EYE2 function in eyespot assembly is redox independent, similar to the auxiliary functions of other thioredoxin family members in protein folding and complex assembly.

39

Dutcher, S. K., W. Gibbons e W. B. Inwood. "A genetic analysis of suppressors of the PF10 mutation in Chlamydomonas reinhardtii." Genetics 120, n. 4 (1 dicembre 1988): 965–76. http://dx.doi.org/10.1093/genetics/120.4.965.

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Abstract A mutation at the PF10 locus of the unicellular green alga Chlamydomonas reinhardtii leads to abnormal cell motility. The asymmetric form of the ciliary beat stroke characteristic of wild-type flagella is modified by this mutation to a nearly symmetric beat. We report here that this abnormal motility is a conditional phenotype that depends on light intensity. In the absence of light or under low light intensities, the motility is more severely impaired than at higher light intensities. By UV mutagenesis we obtained 11 intragenic and 70 extragenic strains that show reversion of the pf10 motility phenotype observed in low light. The intragenic events reverted the motility phenotype of the pf10 mutation completely. The extragenic events define at least seven suppressor loci; these map to linkage groups IV, VII, IX, XI, XII and XVII. Suppressor mutations at two of the seven loci (LIS1 and LIS2) require light for their suppressor activity. Forty-eight of the 70 extragenic suppressors were examined in heterozygous diploid cells; 47 of these mutants were recessive to the wild-type allele and one mutant (bop5-1) was dominant to the wild-type allele. Complementation analysis of the 47 recessive mutants showed unusual patterns. Most mutants within a recombinationally defined group failed to complement one another, although there were pairs that showed intra-allelic complementation. Additionally, some of the mutants at each recombinationally defined locus failed to complement mutants at other loci. They define dominant enhancers of one another.

40

Pandey, Manishi, Gary D. Stormo e Susan K. Dutcher. "Alternative Splicing During the Chlamydomonasreinhardtii Cell Cycle". G3: Genes|Genomes|Genetics 10, n. 10 (18 agosto 2020): 3797–810. http://dx.doi.org/10.1534/g3.120.401622.

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Genome-wide analysis of transcriptome data in Chlamydomonas reinhardtii shows periodic patterns in gene expression levels when cultures are grown under alternating light and dark cycles so that G1 of the cell cycle occurs in the light phase and S/M/G0 occurs during the dark phase. However, alternative splicing, a process that enables a greater protein diversity from a limited set of genes, remains largely unexplored by previous transcriptome based studies in C. reinhardtii. In this study, we used existing longitudinal RNA-seq data obtained during the light-dark cycle to investigate the changes in the alternative splicing pattern and found that 3277 genes (19.75% of 17,746 genes) undergo alternative splicing. These splicing events include Alternative 5′ (Alt 5′), Alternative 3′ (Alt 3′) and Exon skipping (ES) events that are referred as alternative site selection (ASS) events and Intron retention (IR) events. By clustering analysis, we identified a subset of events (26 ASS events and 10 IR events) that show periodic changes in the splicing pattern during the cell cycle. About two-thirds of these 36 genes either introduce a pre-termination codon (PTC) or introduce insertions or deletions into functional domains of the proteins, which implicate splicing in altering gene function. These findings suggest that alternative splicing is also regulated during the Chlamydomonas cell cycle, although not as extensively as changes in gene expression. The longitudinal changes in the alternative splicing pattern during the cell cycle captured by this study provides an important resource to investigate alternative splicing in genes of interest during the cell cycle in Chlamydomonas reinhardtii and other eukaryotes.

41

Bult�, L., e P. Bennoun. "Translational accuracy and sexual differentiation in Chlamydomonas reinhardtii". Current Genetics 18, n. 2 (agosto 1990): 155–60. http://dx.doi.org/10.1007/bf00312603.

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42

Cahoon, A. Bruce, e Ali A. Qureshi. "Leaderless mRNAs are circularized in Chlamydomonas reinhardtii mitochondria". Current Genetics 64, n. 6 (1 giugno 2018): 1321–33. http://dx.doi.org/10.1007/s00294-018-0848-2.

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43

Newman, Scott M., Nicholas W. Gillham, Elizabeth H. Harris, Anita M. Johnson e John E. Boynton. "Targeted disruption of chloroplast genes in Chlamydomonas reinhardtii". Molecular and General Genetics MGG 230, n. 1-2 (novembre 1991): 65–74. http://dx.doi.org/10.1007/bf00290652.

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44

Hall, Leo M., Kenneth B. Taylor e Daniel D. Jones. "Expression of a foreign gene in Chlamydomonas reinhardtii". Gene 124, n. 1 (febbraio 1993): 75–81. http://dx.doi.org/10.1016/0378-1119(93)90763-s.

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45

Pineda, Manuel, Purificaci�n Cabello e Jacobo C�rdenas. "Ammonium regulation of urate uptake in Chlamydomonas reinhardtii". Planta 171, n. 4 (agosto 1987): 496–500. http://dx.doi.org/10.1007/bf00392297.

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46

von Gromoff, Erika D., e Christoph F. Beck. "Genes expressed during sexual differentiation of Chlamydomonas reinhardtii". Molecular and General Genetics MGG 241-241, n. 3-4 (novembre 1993): 415–21. http://dx.doi.org/10.1007/bf00284695.

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47

Cole, Douglas G. "The Intraflagellar Transport Machinery of Chlamydomonas reinhardtii". Traffic 4, n. 7 (9 giugno 2003): 435–42. http://dx.doi.org/10.1034/j.1600-0854.2003.t01-1-00103.x.

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48

Franz, Sophie, Elisabeth Ignatz, Sandra Wenzel, Hannah Zielosko, Eka Putra Gusti Ngurah Putu, Manuel Maestre-Reyna, Ming-Daw Tsai, Junpei Yamamoto, Maria Mittag e Lars-Oliver Essen. "Structure of the bifunctional cryptochrome aCRY from Chlamydomonas reinhardtii". Nucleic Acids Research 46, n. 15 (19 luglio 2018): 8010–22. http://dx.doi.org/10.1093/nar/gky621.

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49

Schnell, R. A., e P. A. Lefebvre. "Isolation of the Chlamydomonas regulatory gene NIT2 by transposon tagging." Genetics 134, n. 3 (1 luglio 1993): 737–47. http://dx.doi.org/10.1093/genetics/134.3.737.

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Abstract Genetic evidence suggests that the NIT2 gene of Chlamydomonas reinhardtii encodes a positive regulator of the nitrate-assimilation pathway. To learn more about the function of the NIT2 gene product, we isolated the gene using a transposon-tagging strategy. A nit2 mutation caused by the insertion of a transposon was identified by testing spontaneous nit2 mutants for the presence of new copies of Gulliver or TOC1, transposable elements that have been identified in Chlamydomonas. In 2 of the 14 different mutants that were analyzed, a Gulliver element was found to be genetically and phenotypically associated with the nit2 mutation. Using the Gulliver element as a probe, one of the transposon-induced nit2 alleles was isolated, and a sequence adjoining the transposon was used to isolate the corresponding wild-type locus. The NIT2 gene was delimited by mapping DNA rearrangements associated with nit2 mutations and mutant rescue by genetic transformation. The NIT2 gene encodes a 6-kb transcript that was not detected in cells grown in the presence of ammonium. Likewise, NIT2-dependent genes are repressed in ammonium-grown cells. These results suggest that repression of the NIT2 gene may mediate metabolite repression of the nitrate assimilation pathway in Chlamydomonas.

50

Dionisio-Sese, Maribel L., Hideya Fukuzawa e Shigetoh Miyachi. "Light-Induced Carbonic Anhydrase Expression in Chlamydomonas reinhardtii". Plant Physiology 94, n. 3 (1 novembre 1990): 1103–10. http://dx.doi.org/10.1104/pp.94.3.1103.

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