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  • 1
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Biochemistry 30 (1991), S. 1127-1131 
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0983
    Keywords: Nicotiana plumbaginifolia ; Chloroplast 16s ribosomal RNA ; Streptomycin resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In a previous paper we reported the isolation of streptomycin-resistant mutants from Nicotiana plumbaginifolia and presented evidence for chloroplast control of the resistance trait. To understand the molecular basis of the resistance in these mutants, we sequenced three regions in the chloroplast 16s rRNA gene, which correspond to the 5′ terminus, the 530 loop, and the 900 stem/loop of Escherichia coli 16s rRNA, and compared them with the sequences of the wild-type. Our results show that: (1) nine mutants have a C to T change at position 912, (2) one mutant (SR1021) has a G to A change at position 885, (3) one mutant has a C to T change at position 526, based on E. coli numbering; and (4) three mutants do not have any change in the regions analyzed. The point mutation detected in SR1021 has not been reported previously. In E. coli 16s rRNA, position 885 is protected from chemical probing by ribosomal protein S12 and is closely juxtaposed with the streptomycin-binding region (positions 912–915) in the predicted secondary structure. It is likely that the G to A transition at this position is a novel mutation for streptomycin resistance.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-5028
    Keywords: Chlamydomonas ; chloroplast ; mRNA ; tufA/y-1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Light induces chloroplast (Cp) differentiation in dark-grown y-1 strains of Chlamydomonas reinhardtii. Slot blot analysis was used to quantitate tufA, psbA, psbK, rbcL, and 16S rRNA transcript accumulation and transcription during Cp differentiation. When etiolated cc-125 y-1 cells were illuminated for 5 h, a 1710 bp tufA mRNA accumulated up to 5-fold while the psbA, rbcL, and 16S rRNA transcripts accumulated less than 1.5-fold. The tufA gene encodes translational elongation factor EF-Tu. The light-induced accumulation of tufA mRNA did not occur in cc-1931, a strain that does not become etiolated in darkness. Pulse labelling was used to measure the transcription of Cp transcripts during tufA mRNA accumulation, and no detectable change in tufA transcription was observed. These results imply that the half life of the tufA transcript increases during the greening process.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 15 (1990), S. 449-455 
    ISSN: 1573-5028
    Keywords: chloroplast ; frx B protein ; spinach ; tobacco ; western blot
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Previously, a ferredoxin-type iron-sulfur protein, frx B protein, was identified in a high-salt extract of the purified thylakoid membrane of Chlamydomonas reinhardtii, a unicellular green alga. Polyclonal antibody was raised against a synthetic pentadecameric peptide with an amino acid sequence corresponding to the highly conserved region of the putative frx B proteins of 3 land plants [21]. In this report, protein(s) reacting strongly and specifically with this antibody was detected in the equivalent high-salt extract prepared from purified chloroplast of spinach and tobacco. One strong reaction polypeptide band from tobacco chloroplast was purified from SDS-polyacrylamide gel and subjected to endoproteinase lys C digestion. The resulting polypeptides were separated by reversed-phase chromatography. N-terminal sequencing of 3 purified polypeptides revealed that the protein is encoded by the ‘frxB gene’ identified from DNA sequence analysis.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 172 (1993), S. 57-63 
    ISSN: 1615-6102
    Keywords: Development ; Fluorescence ; frx B Protein ; Immunocytology ; Membrane potential
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Immunocytology method showed that frx B was primarily located in vesicles adjacent to the pyrenoid inChlamydomonas reinhardtii. In synchronous cells, frx B concentration in these vesicles increased in the light period and decreased in the dark period. Prolonged treatment with 5-fluorodeoxyuridine or aging reduced the detectable cellular frx B content. In these cells, frx B appeared as a single dot attached to the pyrenoid. The frx B content in each mating gamete was initially high. As the mating progressed, frx B-containing vesicles in the male gamete disintegrated while that in the female gamete persisted. The time course of this change coincided with that of the chloroplast DNA nucleoids. Persistant small patches of frx B around the pyrenoid were detected in mature zygotes. Fluorescent membrane potential probe showed that membrane vesicles around the pyrenoid had higher transmembrane potential than other chloroplast membranes. This high membrane potential was sensitive to inhibitors of chlororespiration.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1617-4623
    Keywords: Chloroplast ; Replication origin ; Bent DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We demonstrated that the 1055 by restriction fragment containing OriA, a chloroplast DNA replication origin of Chlamydomonas reinhardtii, has electrophoretic anomalies characteristic of bent DNA. A tandem dimer of the region was constructed. Quantitative measurement of the relative gel mobility of a set of permuted fragments was used to extrapolate the approximate position of the bent DNA segment. By analyzing the gel mobility of short, sequenced fragments of the bent DNA region, the putative bending locus was identified. Two A4 tracts and two A5 tracts were located in the bending locus. Oligonucleotide-directed mutagenesis was then used to disrupt the A tract or the spacing between A tracts and the effect of site-specific mutation on electrophoretic mobility was analyzed. To assess the functional role of the bent DNA region, subclones containing the bending locus, mutated bending locus, and regions flanking the bending locus were constructed. Each subclone was used as template in an in vitro DNA replication system which preferentially initiated DNA replication at OriA. A 224 by subclone with the bending locus positioned in the middle displayed the highest replication function and was sufficient to initiate DNA replication in vitro. Site-specific mutations or alterations of the A tracts resulted in decreased DNA bending and decreased DNA replication activity.
    Type of Medium: Electronic Resource
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