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See detailEpithelial and endothelial expression of the green fluorescent protein reporter gene under the control of bovine prion protein (PrP) gene regulatory sequences in transgenic mice.
Lemaire-Vieille, C.; Schulze, T.; Podevin-Dimster, V. et al

in Proceedings of the National Academy of Sciences of the United States of America (2000), 97(10), 5422-7

The expression of the cellular form of the prion protein (PrP(c)) gene is required for prion replication and neuroinvasion in transmissible spongiform encephalopathies. The identification of the cell ... [more ▼]

The expression of the cellular form of the prion protein (PrP(c)) gene is required for prion replication and neuroinvasion in transmissible spongiform encephalopathies. The identification of the cell types expressing PrP(c) is necessary to understanding how the agent replicates and spreads from peripheral sites to the central nervous system. To determine the nature of the cell types expressing PrP(c), a green fluorescent protein reporter gene was expressed in transgenic mice under the control of 6.9 kb of the bovine PrP gene regulatory sequences. It was shown that the bovine PrP gene is expressed as two populations of mRNA differing by alternative splicing of one 115-bp 5' untranslated exon in 17 different bovine tissues. The analysis of transgenic mice showed reporter gene expression in some cells that have been identified as expressing PrP, such as cerebellar Purkinje cells, lymphocytes, and keratinocytes. In addition, expression of green fluorescent protein was observed in the plexus of the enteric nervous system and in a restricted subset of cells not yet clearly identified as expressing PrP: the epithelial cells of the thymic medullary and the endothelial cells of both the mucosal capillaries of the intestine and the renal capillaries. These data provide valuable information on the distribution of PrP(c) at the cellular level and argue for roles of the epithelial and endothelial cells in the spread of infection from the periphery to the brain. Moreover, the transgenic mice described in this paper provide a model that will allow for the study of the transcriptional activity of the PrP gene promoter in response to scrapie infection. [less ▲]

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See detailOpposing Actions of Intact and N-Terminal Fragments of the Human Prolactin/Growth Hormone Family Members on Angiogenesis: An Efficient Mechanism for the Regulation of Angiogenesis
Struman, Ingrid ULg; Bentzien, F.; Lee, H. et al

in Proceedings of the National Academy of Sciences of the United States of America (1999), 96(4), 1246-51

Angiogenesis, the process of development of a new microvasculature, is regulated by a balance of positive and negative factors. We show both in vivo and in vitro that the members of the human prolactin ... [more ▼]

Angiogenesis, the process of development of a new microvasculature, is regulated by a balance of positive and negative factors. We show both in vivo and in vitro that the members of the human prolactin/growth hormone family, i.e., human prolactin, human growth hormone, human placental lactogen, and human growth hormone variant are angiogenic whereas their respective 16-kDa N-terminal fragments are antiangiogenic. The opposite actions are regulated in part via activation or inhibition of mitogen-activated protein kinase signaling pathway. In addition, the N-terminal fragments stimulate expression of type 1 plasminogen activator inhibitor whereas the intact molecules have no effect, an observation consistent with the fragments acting via separate receptors. The concept that a single molecule encodes both angiogenic and antiangiogenic peptides represents an efficient model for regulating the balance of positive and negative factors controlling angiogenesis. This hypothesis has potential physiological importance for the control of the vascular connection between the fetal and maternal circulations in the placenta, where human prolactin, human placental lactogen, and human growth hormone variant are expressed. [less ▲]

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See detailFine-Mapping of Quantitative Trait Loci by Identity by Descent in Outbred Populations: Application to Milk Production in Dairy Cattle
Riquet, J.; Coppieters, Wouter ULg; Cambisano, Nadine ULg et al

in Proceedings of the National Academy of Sciences of the United States of America (1999), 96(16), 9252-9257

We previously mapped a quantitative trait locus (QTL) affecting milk production to bovine chromosome 14. To refine the map position of this QTL, we have increased the density of the genetic map of ... [more ▼]

We previously mapped a quantitative trait locus (QTL) affecting milk production to bovine chromosome 14. To refine the map position of this QTL, we have increased the density of the genetic map of BTA14q11-16 by addition of nine microsatellites and three single nucleotide polymorphisms. Fine-mapping of the QTL was accomplished by a two-tiered approach. In the first phase, we identified seven sires heterozygous "Qq" for the QTL by marker-assisted segregation analysis in a Holstein-Friesian pedigree comprising 1,158 individuals. In a second phase, we genotyped the seven selected sires for the newly developed high-density marker map and searched for a shared haplotype flanking an hypothetical, identical-by-descent QTL allele with large substitution effect. The seven chromosomes increasing milk fat percentage were indeed shown to carry a common chromosome segment with an estimated size of 5 cM predicted to contain the studied QTL. The same haplotype was shown to be associated with increased fat percentage in the general population as well, providing additional support in favor of the location of the QTL within the corresponding interval. [less ▲]

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See detailImpaired motor coordination and Purkinje cell excitability in mice lacking calretinin
Schiffmann, Serge N.; Cheron, Guy; Lohoff, Ann et al

in Proceedings of the National Academy of Sciences of the United States of America (1999), 96

n the cerebellum, the parallel fiber-Purkinje cell synapse can undergo long-term synaptic plasticity suggested to underlie motor learning and resulting from variations in intracellular calcium ... [more ▼]

n the cerebellum, the parallel fiber-Purkinje cell synapse can undergo long-term synaptic plasticity suggested to underlie motor learning and resulting from variations in intracellular calcium concentration ([Ca2+]i). Ca2+ binding proteins are enriched in the cerebellum, but their role in information processing is not clear. Here, we show that mice deficient in calretinin (Cr-/-) are impaired in tests of motor coordination. An impairment in Ca2+ homeostasis in Cr-/- Purkinje cells was supported by the high Ca2+-saturation of calbindin-D28k in these cells. The firing behavior of Purkinje cells is severely affected in Cr-/- alert mice, with alterations of simple spike firing rate, complex spike duration, and simple spike pause. In contrast, in slices, transmission at parallel fiber- or climbing fiber-Purkinje cell synapses is unaltered, indicating that marked modifications of the firing behavior in vivo can be undetectable in slice. Thus, these results show that calretinin plays a major role at the network level in cerebellar physiology [less ▲]

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See detailExtracellular enveloped vaccinia virus is resistant to complement because of incorporation of host complement control proteins into its envelope
Vanderplasschen, Alain ULg; Mathew, E.; Hollinshead, M. et al

in Proceedings of the National Academy of Sciences of the United States of America (1998), 95

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See detailcDNA cloning and expression of bovine procollagen I N-proteinase: a new member of the superfamily of zinc-metalloproteinases with binding sites for cells and other matrix components.
Colige, Alain ULg; Li, S. W.; Sieron, A. L. et al

in Proceedings of the National Academy of Sciences of the United States of America (1997), 94(6), 2374-9

Procollagen N-proteinase (EC 3.4.24.14) cleaves the amino-propeptides in the processing of type I and type II procollagens to collagens. Deficiencies of the enzyme cause dermatosparaxis in cattle and ... [more ▼]

Procollagen N-proteinase (EC 3.4.24.14) cleaves the amino-propeptides in the processing of type I and type II procollagens to collagens. Deficiencies of the enzyme cause dermatosparaxis in cattle and sheep, and they cause type VIIC Ehlers-Danlos syndrome in humans, heritable disorders characterized by accumulation of pNcollagen and severe skin fragility. Amino acid sequences for the N-proteinase were used to obtain cDNAs from bovine skin. Three overlapping cDNAs had an ORF coding for a protein of 1205 residues. Mammalian cells stably transfected with a complete cDNA secreted an active recombinant enzyme that specifically cleaved type I procollagen. The protein contained zinc-binding sequences of the clan MB of metallopeptidases that includes procollagen C-proteinase/BMP-1. The protein also contained four repeats that are homologous to domains found in thrombospondins and in properdin and that can participate in complex intermolecular interactions such as activation of latent forms of transforming growth factor beta or the binding to sulfatides. Therefore, the enzyme may play a role in development that is independent of its role in collagen biosynthesis. This hypothesis was supported by the observation that in some tissues the levels of mRNA for the enzyme are disproportionately high relative to the apparent rate of collagen biosynthesis. [less ▲]

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See detailImpaired LTP induction in dentate gyrus of calretinin-deficient mice
Schurmans, Stéphane ULg; Schiffmann, S. N.; Gurden, H. et al

in Proceedings of the National Academy of Sciences of the United States of America (1997), 94

Calretinin (Cr) is a Ca21 binding protein present in various populations of neurons distributed in the central and peripheral nervous systems. We have generated Cr-deficient (Cr2/2) mice by gene targeting ... [more ▼]

Calretinin (Cr) is a Ca21 binding protein present in various populations of neurons distributed in the central and peripheral nervous systems. We have generated Cr-deficient (Cr2/2) mice by gene targeting and have investigated the associated phenotype. Cr2/2 mice were viable, and a large number of morphological, biochemical, and behavioral parameters were found unaffected. In the normal mouse hippocampus, Cr is expressed in a widely distributed subset of GABAergic interneurons and in hilar mossy cells of the dentate gyrus. Because both types of cells are part of local pathways innervating dentate granule cells andyor pyramidal neurons, we have explored in Cr2/2 mice the synaptic transmission between the perforant pathway and granule cells and at the Schaffer commissural input to CA1 pyramidal neurons. Cr2/2 mice showed no alteration in basal synaptic transmission, but long-term potentiation (LTP) was impaired in the dentate gyrus. Normal LTP could be restored in the presence of the GABAA receptor antagonist bicuculline, suggesting that in Cr2/2 dentate gyrus an excess of g-minobutyric acid (GABA) release interferes with LTP induction. Synaptic transmission and LTP were normal in CA1 area, which contains only few Cr-positive GABAergic interneurons. Cr2/2 mice performed normally in spatial memory task. These results suggest that expression of Cr contributes to the control of synaptic plasticity in mouse dentate gyrus by indirectly regulating the activity of GABAergic interneurons, and that Cr2/2 mice represent a useful tool to understand the role of dentate LTP in learning and memory [less ▲]

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See detailA novel seizure-induced synaptotagmin gene identified by differential display.
Babity, J. M.; Armstrong, J. N.; Plumier, Jean-Christophe ULg et al

in Proceedings of the National Academy of Sciences of the United States of America (1997), 94(6), 2638-41

Systemic administration of kainic acid, a cyclic analogue of glutamate, produces many of the clinical features of human temporal lobe epilepsy and status epilepticus in rats, including the induction of ... [more ▼]

Systemic administration of kainic acid, a cyclic analogue of glutamate, produces many of the clinical features of human temporal lobe epilepsy and status epilepticus in rats, including the induction of motor convulsions and the degeneration of neurons in the hippocampus and piriform cortex. Differential display PCR was used to identify mRNAs that are differentially expressed between degenerating and nondegenerating tissues in the brain after kainic acid-induced seizure activity. A novel cDNA fragment expressed in the degenerating hippocampus and piriform cortex, but not in the nondegenerating parietal cortex, was identified, cloned, and sequenced. This novel cDNA fragment identified a new member of the synaptotagmin gene family that is rapidly and transiently induced in response to seizure activity. Differential expression of this synaptotagmin gene, syt X, was confirmed by Northern blot analysis and in situ hybridization. This novel, inducible synaptotagmin gene may provide a direct link between seizure-induced neuronal gene expression and subsequent modulation of synaptic structure and function. [less ▲]

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See detailTranscription factor TFIIH and DNA endonuclease Rad2 constitute yeast nucleotide excision repair factor 3: implications for nucleotide excision repair and Cockayne syndrome
Habraken, Yvette ULg; Sung, Patrick; Prakash, Louise et al

in Proceedings of the National Academy of Sciences of the United States of America (1996), 93(20), 10718-22

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See detailExpression of protein encoded by varicella-zoster virus open reading frame 63 in latently infected human ganglionic neurons
Mahalingam, Ravy; Wellish, Mary; Cohrs, Randall et al

in Proceedings of the National Academy of Sciences of the United States of America (1996), 93(5), 2122-2124

The ganglionic cell type in which varicellazoster virus (VZV) is latent in humans was analyzed by using antibodies raised against in vitro-expressed VZV open reading frame 63 protein, VZV open reading ... [more ▼]

The ganglionic cell type in which varicellazoster virus (VZV) is latent in humans was analyzed by using antibodies raised against in vitro-expressed VZV open reading frame 63 protein, VZV open reading frame 63 protein was detected exclusively in the cytoplasm of neurons of latently infected human trigeminal and thoracic ganglia. This is, to our knowledge, the first identification of a herpesvirus protein expressed during latency in the human nervous system. [less ▲]

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See detailThe catalytic mechanism of beta-lactamases: NMR titration of an active-site lysine residue of the TEM-1 enzyme.
Damblon, Christian ULg; Raquet, X.; Lian, L. Y. et al

in Proceedings of the National Academy of Sciences of the United States of America (1996), 93(5), 1747-52

Beta-Lactamases are widespread in the bacterial world, where they are responsible for resistance to penicillins, cephalosporins, and related compounds, currently the most widely used antibacterial agents ... [more ▼]

Beta-Lactamases are widespread in the bacterial world, where they are responsible for resistance to penicillins, cephalosporins, and related compounds, currently the most widely used antibacterial agents. Detailed structural and mechanistic understanding of these enzymes can be expected to guide the design of new antibacterial compounds resistant to their action. A number of high-resolution structures are available for class A beta-lactamases, whose catalytic mechanism involves the acylation of a serine residue at the active site. The identity of the general base which participates in the activation of this serine residue during catalysis has been the subject of controversy, both a lysine residue and a glutamic acid residue having been proposed as candidates for this role. We have used the pH dependence of chemical modification of epsilon-amino groups by 2,4,6,-trinitrobenzenesulfonate and the pH dependence of the epsilon-methylene 1H and 13C chemical shifts (in enzyme selectively labeled with [epsilon-13C]lysine) to estimate the pKa of the relevant lysine residue, lysine-73, of TEM-1 beta-lactamase. Both methods show that the pKa of this residue is > 10, making it very unlikely that this residue could act as a proton acceptor in catalysis. An alternative mechanism in which this role is performed by glutamate-166 through an intervening water molecule is described. [less ▲]

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See detailEmergence of the ZNF91 Kruppel-associated box-containing zinc finger gene family in the last common ancestor of anthropoidea
Bellefroid, Eric J.; Marine, Jean-Christophe; Matera, A Gregory et al

in Proceedings of the National Academy of Sciences of the United States of America (1995), 92(23), 10757-61

The ZNF91 gene family, a subset of the Kruppel-associated box (KRAB)-containing group of zinc finger genes, comprises more than 40 loci; most reside on human chromosome 19p12-p13.1. We have examined the ... [more ▼]

The ZNF91 gene family, a subset of the Kruppel-associated box (KRAB)-containing group of zinc finger genes, comprises more than 40 loci; most reside on human chromosome 19p12-p13.1. We have examined the emergence and evolutionary conservation of the ZNF91 family. ZNF91 family members were detected in all species of great apes, gibbons, Old World monkeys, and New World monkeys examined but were not found in prosimians or rodents. In each species containing the ZNF91 family, the genes were clustered at one major site, on the chromosome(s) syntenic to human chromosome 19. To identify a putative "founder" gene, > 20 murine KRAB-containing zinc finger protein (ZFP) cDNAs were randomly cloned, but none showed sequence similarity to the ZNF91 genes. These observations suggest that the ZNF91 gene cluster is a derived character specific to Anthropoidea, resulting from a duplication and amplification event some 55 million years ago in the common ancestor of simians. Although the ZNF91 gene cluster is present in all simian species, the sequences of the human ZNF91 gene that confer DNA-binding specificity were conserved only in great apes, suggesting that there is not a high selective pressure to maintain the DNA targets of these proteins during evolution. [less ▲]

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See detailActivation of mitogen-activated protein kinases by vascular endothelial growth factor and basic fibroblast growth factor in capillary endothelial cells is inhibited by the antiangiogenic factor 16-kDa N-terminal fragment of prolactin
D'Angelo, Gisela; Struman, Ingrid ULg; Martial, Joseph ULg et al

in Proceedings of the National Academy of Sciences of the United States of America (1995), 92(14), 6374-8

A number of factors both stimulating and inhibiting angiogenesis have been described. In the current work, we demonstrate that the angiogenic factor vascular endothelial growth factor (VEGF) activates ... [more ▼]

A number of factors both stimulating and inhibiting angiogenesis have been described. In the current work, we demonstrate that the angiogenic factor vascular endothelial growth factor (VEGF) activates mitogen-activated protein kinase (MAPK) as has been previously shown for basic fibroblast growth factor. The antiagiogenic factor 16-kDa N-terminal fragment of human prolactin inhibits activation of MAPK distal to autophosphorylation of the putative VEGF receptor, Flk-1, and phospholipase C-gamma. These data show that activation and inhibition of MAPK may play a central role in the control of angiogenesis. [less ▲]

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See detailAttenuation Of Bovine Leukemia-Virus By Deletion Of R3 And G4 Open Reading Frames
Willems, Luc ULg; Kerkhofs, P.; Dequiedt, Franck ULg et al

in Proceedings of the National Academy of Sciences of the United States of America (1994), 91(24),

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See detailChromosomal localization of the callipyge gene in sheep (Ovis aries) using bovine DNA markers
Cockett, N. E.; Jackson, S. P.; Shay, T. D. et al

in Proceedings of the National Academy of Sciences of the United States of America (1994), 91

A mutation causing muscular hypertrophy, with associated leanness and improved feed efficiency, has been recently identified in domestic sheep (Ovis aries). Preliminary results indicate that an autosomal ... [more ▼]

A mutation causing muscular hypertrophy, with associated leanness and improved feed efficiency, has been recently identified in domestic sheep (Ovis aries). Preliminary results indicate that an autosomal dominant gene may be responsible for this economically advantageous trait. We have exploited the conservation in sequence and chromosomal location of DNA markers across Bovidae to map the corresponding callipyge locus to ovine chromosome 18 using a battery of bovine chromosome 21 markers. Chromosomal localization of the ovine callipyge locus is the first step toward positional cloning of the corresponding gene. [less ▲]

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See detailMicrosatellite mapping of the gene causing weaver disease in cattle will allow the study of an associated QTL
Georges, Michel ULg; Lathrop, M.; Dietz, A. B. et al

in Proceedings of the National Academy of Sciences of the United States of America (1993), 90

A genetic disease in cattle, progressive degenerative myeloencephalopathy (weaver disease), is associated with increased milk production. This association could result from population stratification, from ... [more ▼]

A genetic disease in cattle, progressive degenerative myeloencephalopathy (weaver disease), is associated with increased milk production. This association could result from population stratification, from a pleiotropic effect of a single gene, or from linkage disequilibrium between the gene causing weaver disease and a quantitative trait locus (QTL) for milk production. To test these hypotheses, we performed an extensive linkage study in a bovine pedigree segregating for the weaver condition and identified a microsatellite locus (TGLA116) closely linked to the weaver gene (zmax, 8.15; theta, 0.03). TGLA116 and, by extension, the weaver locus were assigned to bovine synteny group 13. This microsatellite can be used to identify weaver carriers, to select against this genetic defect, and to study the effect of the corresponding chromosomal region on milk production in Brown Swiss and other breeds of cattle. [less ▲]

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See detailMutations In The Bovine Leukemia-Virus Tax Protein Can Abrogate The Long Terminal Repeat-Directed Transactivating Activity Without Concomitant Loss Of Transforming Potential
Willems, Luc ULg; Grimonpont, C.; Heremans, H. et al

in Proceedings of the National Academy of Sciences of the United States of America (1992), 89(9),

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See detailFusogenic Segments Of Bovine Leukemia-Virus And Simian Immunodeficiency Virus Are Interchangeable And Mediate Fusion By Means Of Oblique Insertion In The Lipid Bilayer Of Their Target-Cells
Voneche, V.; Portetelle, Daniel ULg; Kettmann, Richard ULg et al

in Proceedings of the National Academy of Sciences of the United States of America (1992), 89(9),

Modified bovine leukemia virus (BLV) glycoproteins were expressed by using vaccinia virus recombinants, and their fusogenic capacities were examined by a syncytia-formation assay. This analysis indicates ... [more ▼]

Modified bovine leukemia virus (BLV) glycoproteins were expressed by using vaccinia virus recombinants, and their fusogenic capacities were examined by a syncytia-formation assay. This analysis indicates that (i) both BLV envelope glycoproteins gp51 and gp30 are necessary for cell fusion; (ii) insertion of the N-terminal segment of gp30 (fusion peptide) into the lipid bilayer in an oblique orientation, as predicted by computer conformational analysis, results in fusogenic capacities higher than insertion in a perpendicular or parallel orientation; and (iii) replacement of the BLV fusion peptide with its simian immunodeficiency virus counterpart does not modify the fusogenic capacity of the BLV glycoprotein. [less ▲]

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See detailIdentification of the major pregnancy-specific antigens of cattle and sheep as inactive members of the aspartic proteinase family
Xie, Sancai; Low, Boon G.; Nagel, Robert J. et al

in Proceedings of the National Academy of Sciences of the United States of America (1991), 88(22), 10247-10251

Pregnancy in cattle and sheep can be diagnosed by the presence of a conceptus-derived antigen in maternal serum that is secreted by trophoblast and placental tissue primarily as an acidic component of Mr ... [more ▼]

Pregnancy in cattle and sheep can be diagnosed by the presence of a conceptus-derived antigen in maternal serum that is secreted by trophoblast and placental tissue primarily as an acidic component of Mr 67,000. Molecular cloning of its cDNA reveals that the antigen belongs to the aspartic proteinase family and has greater than 50% amino acid sequence identity to pepsin, cathepsin D, and cathepsin E. The inferred sequences of the ovine and bovine polypeptides show approximately 73% identity to each other. Critical amino acid substitutions at the active site regions suggest that both proteins are enzymatically inactive. The antigen is a product of trophoblast binucleate cells that invade maternal endometrium at implantation sites. [less ▲]

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See detailCYTODUCTION IN CHLAMYDOMONAS-REINHARDTII
Matagne, René-Fernand ULg; Remacle, Claire ULg; DINANT, M.

in Proceedings of the National Academy of Sciences of the United States of America (1991), 88(16), 7447-7450

After conjugation between Chlamydomonas gametes of opposite mating type, a transient dikaryon is formed. The two nuclei fuse within 4-6 hr after mating. The young diploid zygote differentiates into ... [more ▼]

After conjugation between Chlamydomonas gametes of opposite mating type, a transient dikaryon is formed. The two nuclei fuse within 4-6 hr after mating. The young diploid zygote differentiates into dormant zygospore competent to complete meiosis, or more rarely (2-10% of cases) it undergoes mitosis to produce a stable diploid progeny. We here bring genetical, biochemical, and cytological evidence that among the mitotic zygotes, a large proportion of them undergo cytokinesis without fusion of the nuclei - a process that has been termed "cytoduction." By using appropriate genetic markers, haploid cytoductants that possess the nuclear genotype of one parent and the chloroplast marker of the other parent can easily be isolated. Genetical analysis and hybridization experiments moreover show that many haploid cytoductants transmit the chloroplast DNA molecules of both parents and that, as in diploids, these DNA copies occasionally recombine. This process of cytoduction extends the life cycle of Chlamydomonas and provides new tools for its genetic analysis. [less ▲]

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