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  • 1
    ISSN: 1432-1432
    Keywords: Key words: Brown alga —cox1 gene — Group IIA and IIB introns — Phylogeny — Reverse transcriptase-like genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. The mitochondrial genome of the brown alga Pylaiella littoralis contains two different types of group II introns. They each encode complete complex proteins, i.e., with a reverse transcriptase domain, a maturase or X domain, and an endonuclease or H-N-H/zinc finger domain. To our knowledge, this is the first example of the presence in the same genome of introns belonging to subgroups IIA and IIB which both contain multidomained RT-like proteins. We describe the group IIA introns that interrupt the cox1 gene. The RT-like proteins contained in these introns were compared to those of the LSU rDNA group IIB introns. The phylogenetic relationships of these intron ORFs were investigated and the possible evolution of group II introns is discussed.
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  • 2
    ISSN: 1432-1432
    Keywords: Molecular phylogeny of plastids and mitochondria ; Endosymbiotic origin ; Monophylesis ; Cyanobacteria ; Purple bacteria ; Transit peptide ; Neighbor joining method ; Bootstrapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Algae are a heterogeneous group of photosynthetic eukaryotes traditionally separated into three major subdivisions: rhodophytes, chlorophytes, and chromophytes. The evolutionary origin of rhodophytes or red algae and their links to other photosynthetic and nonphotosynthetic eukaryotes have been a matter of much controversy and speculation. Here we present the first cDNAs of nuclear protein genes from red algae: Those encoding cytosolic and chloroplast glyceraldehyde-3-phosphate dehydrogenases (GAPDH) from Chondrus crispus. A phylogenetic analysis including GAPDH gene sequences from a number of eukaryotic taxa, cyanobacteria, and purple bacteria suggests that chloroplasts and rhodoplasts together form a monophyletic group of cyanobacterial descent and that rhodophytes separated from chlorophytes at about the same time as animals and fungi. The composite GAPDH tree further demonstrates that chloroplast and cytosolic GAPDH genes are closely related to their homologs in cyanobacteria and purple bacteria, respectively, the presumptive ancestors of chloroplasts and mitochondria, thereby firmly establishing the endosymbiotic origin of these nuclear genes and their fixation in eukaryotic cells before the rhodophyte/chlorophyte separation. The present data are in conflict with phylogenetic inferences based on plastid-encoded rbcL sequences supporting a polyphyletic origin of rhodoplasts and chloroplasts. Comparison of rbcL to GAPDH phylogenies suggests that rbcL trees may be misleading because they are composed of branches representing ancient duplicated (paralogous) genes.
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  • 3
    Electronic Resource
    Electronic Resource
    Copenhagen : International Union of Crystallography (IUCr)
    Acta crystallographica 56 (2000), S. 766-768 
    ISSN: 1399-0047
    Source: Crystallography Journals Online : IUCR Backfile Archive 1948-2001
    Topics: Chemistry and Pharmacology , Geosciences , Physics
    Notes: This is the first crystallization report of a glycoside hydrolase which belongs to family 82. A recombinant form of His-tagged ι-carrageenase from Alteromonas fortis was expressed, purified and crystallized. Crystals were obtained by the vapour-diffusion method using polyethylene glycol (MW = 6000) as a precipitant. They belong to space group P21, with unit-cell parameters a = 56.75, b = 91.04, c = 125.01 Å, β = 93.41°. The unit cell contains two molecules in the asymmetric unit related by a non-crystallographic twofold axis. Crystals diffracted to 2.0 Å resolution on a synchrotron beamline.
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  • 4
    ISSN: 1399-0047
    Source: Crystallography Journals Online : IUCR Backfile Archive 1948-2001
    Topics: Chemistry and Pharmacology , Geosciences , Physics
    Notes: A recombinant form of His-tagged κ-carrageenase from Pseudoalteromonas carrageenovora has been expressed, purified and crystallized. Crystals have been obtained by the vapour-diffusion method using polyethylene glycol (Mr = 4000) as a precipitant. These crystals belong to the space group P212121, with unit-cell parameters a  =  58.2, b  =  62.8, c  =  77.9 Å, and diffract to 2.2 Å resolution on a rotating-anode X-ray source.
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  • 5
    ISSN: 1432-0983
    Keywords: Chondrus crispus ; Actin ; Promoter region Intron
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have characterized the cDNA and genomic sequences that encode actin from the multicellular red alga Chondrus crispus. Southern-blot analysis indicates that the C. crispus actin gene (ChAc) is present as a single copy. Northern analysis shows that, like the GapA gene, the actin gene is well expressed in gametophytes but weakly in protoplasts. Compared to actin genes of animals, fungi, green plants and oomycetes, that of C. crispus displays a higher evolutionary rate and does not show any of the amino-acid signatures characteristic of the other lineages. As previously described for GapA, ChAc is interrupted by a single intron at the beginning of the coding region. The site of initiation of transcription was characterized by RNAse protection. The promoter region displays a CAAT box but lacks a canonical TATA motif. Other noticeable features, such as a high content of pyrimidines as well as a 14-nt motif found in both the 5′-untranslated region and the intron, were observed.
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  • 6
    ISSN: 1573-5028
    Keywords: ACEDB ; EST ; Kozak sequence ; Laminaria digitata ; 3′-UTR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Laminariales (Phaeophyceae, Heterokonta) are characterised by a heteromorphic digenetic life cycle with a filamentous, microscopic gametophyte and a highly evolved, macroscopic sporophyte. With the ultimate goal of comparing gene expression in each life cycle stage, complementary DNA libraries were constructed from sporophytes and gametophytes of Laminaria digitata. A set of ca. 500 expressed sequence tags (EST) was generated from each life history phase, by single-run partial sequencing of randomly picked cDNA clones. Comparison of the EST deduced amino acid sequences with database protein sequences assigned a putative identity for 39% of the 412 gametophyte clones and 48% of the 493 sporophyte clones sequenced thus far. These data represent more than 152 different proteins now probably identified in L. digitata. Several of those newly identified proteins are of interest to our understanding of the molecular physiology of kelps, for example their carbon-concentrating mechanisms, cell wall biosynthesis and halogen metabolism. EST analysis also confirmed that genes with long 3′-UTRs are widespread in Laminariales and the study of 5′-UTRs allowed the identification of a Kozak consensus sequence, c(A/C)A(A/C)CAUGGc(G/T). Several potential developmentally regulated differences in gene expression are discussed.
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  • 7
    ISSN: 1573-5028
    Keywords: chloroplast evolution ; chromophyte algae ; 23S rRNA genes ; secondary structure ; nucleotide sequence ; endosymbiosis ; phylogenetic tree
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The entire nucleotide sequence of a 23S rRNA gene from the brown alga Pylaiella littoralis (L.) Kjellm has been determined. The predicted length of the 23S rRNA is 2948 nucleotides, including the 4.5S rRNA-like region at the 3′ end of the molecule. The putative transcript has been folded into a secondary structure by comparison to existing structure models, and the predicted helical regions were inspected by identifying compensatory downstream base changes. The 23S rRNA secondary structure presented here has features that are unique to P. littoralis (no other chromophyte or red algal 23S rRNA sequences are yet available), but has none of the features specific to the chloroplast rRNAs of green plants and green algae. The Pylaiella sequence was aligned with analogous plastidial and eubacterial gene sequences, and the alignment was used to construct a phylogenetic tree. The plastidial sequences formed a coherent cluster closely associated with the 23S rRNA of the cyanobacterium Anacystis nidulans. Within the plastid group, the P. littoralis sequence was most closely related to that of Euglena gracilis confirming earlier analyses based upon 16S rRNA sequences.
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  • 8
    ISSN: 1573-5028
    Keywords: cis-acting elements ; intron conservation ; intron secondary structure ; pre-mRNA splicing ; CpG suppression ; protoplasts ; transcript levels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Our previous phylogenetic analysis based on cDNA sequences of chloroplast and cytosolic glyceraldehyde-3-phosphate dehydrogenases (GAPDH; genes GapA and GapC, respectively) of the red alga Chondrus crispus suggested that rhodophytes and green plants are sister groups with respect to plastids and mitochondria and diverged at about the same time or somewhat later than animals and fungi. Here we characterize the genomic sequences of genes GapC and GapA of C. crispus with respect to promotor structures, intron/exon organization, genomic complexity, G+C content, CpG suppression and their transcript levels in gametophytes and protoplasts, respectively. To our knowledge this is the first report on nuclear protein genes of red algae. The GapC gene is G+C-rich, contains no introns and displays a number of classic sequence motifs within its promotor region, such as TATA, CAAT, GC boxes and several elements resembling the plant-specific G-box palindrome. The GapA gene has a moderate G+C content, a single CAAT box motif in its promotor region and a single intron of 115 bp near its 5′ end. This intron occupies a conserved position corresponding to that of intron 1 in the transit peptide region of chloroplast GAPDH genes (GapA and GapB) of higher plants. It has consensus sequences similar to those of yeast introns and folds into a conspicuous secondary structure of - 61.3 kJ. CpG profiles of genes GapC and GapA and their flanking sequences show no significant CpG depletion suggesting that these genomic sequences are not methylated. Genomic Southern blots hybridized with generic and gene specific probes indicate that both genes are encoded by single loci composed of multiple polymorphic alleles. Northern hybridizations demonstrate that both genes are expressed in gametophytes but not in protoplasts where appreciable amounts of transcripts can only be detected for GapC.
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  • 9
    ISSN: 1573-5028
    Keywords: Chondrus crispus ; mitochondrial genome ; gene map ; restriction map ; rhodophytes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Organellar DNA, i.e. a mixture of plastid and mitochondrial DNAs, was purified from the rhodophyte Chondrus crispus and analysed with restriction endonucleases. Mitochondrial DNA fragments were identified by heterologous hybridization, cloned, mapped and partially sequenced. The mitochondrial genome of C. crispus consists of a 25.9 kb circular molecule on which twenty genes were localized. Compared with other plant mitochondrial genomes, C. crispus mitochondrial DNA appears as a relatively small molecule with a high coding capacity and a specific gene organization. The use of a modified genetic code and the absence of RNA editing, previously reported for the cox3 gene, is a general characteristic of the sequenced genes of this molecule. This is the first detailed description of a red algal mitochondrial genome.
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  • 10
    ISSN: 1573-5176
    Keywords: Delesseria sanguinea ; cultivation ; anticoagulant ; cell-wall ; polysaccharides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tank cultivation ofDelesseria sanguinea was investigated in order to manipulate conditions for vegetative growth and to provide biomass for the analysis of cell wall polysaccharides. Seasonality is subject to short-day photoperiodic control. Night-break or long-day conditions prevented fertility in tetrasporophytes and gametophytes and triggered outgrowth of new blades. Long-day illuminations allowed a 1% daily growth rate. Seawater temperature below 13 °C was necessary for inducing formation of new blades. Both wild and cultivated ofD. sanguinea plants contained a non gelling sulfated heteropolysaccharide composed of a galactosyl backbone branched with xylosyl residues. The hot water extract at neutral pH displayed the highest anticoagulant activity (5 μg ml-1 polysaccharide concentration in APTT clotting assay). No obvious differences were found in polysaccharide chemical composition and properties between gametophytes and sporophytes or between cultivated and wild plants.
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