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  • 1
    Electronic Resource
    Electronic Resource
    Demonstration of in vitro starch branching enzyme activity for a 51/50-kDa polypeptide isolated from developing barley (Hordeum vulgare) caryopses (1996)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 96 (1996), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Starch branching enzyme (SBE, EC 2.4.1.18) activity was followed in developing barley (Hordeum vulgare L. cv. Golf) caryopses during a period of one month after anthesis. Caryopses with the highest specific activity, and corresponding to a fresh weight of around 60 mg per caryopsis, were homogenized and the soluble extract used for branching enzyme purification by FPLC chromatography. Four branching enzyme activity fractions were resolved. From one of these fractions, which exhibited high activity in both the phosphorylation stimulation and amylose branching assays, a branching enzyme preparation containing two related polypeptides of 51 and 50 kDa was obtained. Native polyacrylamide gel electrophoresis and gel filtration showed that the 51/50-kDa polypeptide is monomeric. A combination of phosphorylation stimulation and amylose branching gel assays, SDS-PAGE, and TLC was used to demonstrate the branching activity of the 51/50-kDa polypeptide. The activity was further confirmed by spectroscopic analyses of iodine-glucan complexes. SBEs from four different plant species were compared using the phosphorylation stimulation gel assay.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1996.tb00461.x
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  • 2
    Electronic Resource
    Electronic Resource
    Green fluorescent protein as a reporter system in the transformation of barley cultivars (1999)
    Copenhagen : Munksgaard International Publishers
    Physiologia plantarum 107 (1999), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A cereal transformation vector, pN1473, containing the strong constitutive rice actin promoter Act-1, a multiple cloning site, and the nos terminator, was constructed. Fusion of a plant-optimized gfp gene to Act-1 in pN1473 resulted in the vector pN1473GFP. To assess the suitability of pN1473, and GFP as a reporter system in barley transformation, two barley cultivars (Baronesse and Golden Promise) were transformed by microprojectile bombardment. Transient gfp expression in transformed embryogenic callus material was detectable by fluorescence microscopy less than 12 h after transformation. The presence of the gfp gene in callus and regenerated plantlets was confirmed by PCR amplification and DNA gel-blot analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1034/j.1399-3054.1999.100207.x
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  • 3
    Electronic Resource
    Electronic Resource
    Changes of amino acid sequence in PEST-like area and QEEET motif affect degradation rate of D1 polypeptide in photosystem II (1994)
    Springer
    Plant molecular biology 25 (1994), S. 517-526 
    Details
    ISSN: 1573-5028
    Keywords: D1 polypeptide ; photoinhibition ; photosystem II ; protein turnover ; site-specific mutants ; Synechocystis 6803
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The degradation rate of the D1 polypeptide was measured in threeSynechocystis PCC 6803 mutantsin vivo. Mutations were introduced into a putative cleavage area of the D1 polypeptide (QEEET motif) and into the PEST-like area. PEST sequences are often found in proteins with a high turnover rate. The QEEET-motif mutants are CA1 [Δ(E242-E244);Q241H] and E243K, and the third mutation, E229D, was directed to the PEST-like area. During high-light illumination (1500 μmol photons m-2s-1) that induced photoinhibition of photosystem II (PSII), the half-life time of the D1 polypeptide in mutant E229D (t 1/2=35 min) was about twice as long as in AR (control strain) cells (t 1/2=19 min). In growth light (40 μmol photons m-2s-1), the degradation rate of the D1 polypeptide in E229D and AR strains was the same (t 1/2≈5 h). In growth light the D1 polypeptide was degraded faster in both QEEET-motif mutants than in the AR strain, but in photoinhibitory light the degradation rates were similar. According to these results, the highly conservative QEEET motif as such is not required for the proteolytic cut of the D1 polypeptide, but it does affect the rate of degradation. No simple correlation existed between the degradation rate of the D1 polypeptide and the susceptibility of PSII to photoinhibition in mutant and AR cells under our experimental conditions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00043879
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  • 4
    Electronic Resource
    Electronic Resource
    Influence of light on accumulation of photosynthesis-specific transcripts in the cyanobacterium Synechocystis 6803 (1989)
    Springer
    Plant molecular biology 13 (1989), S. 693-700 
    Details
    ISSN: 1573-5028
    Keywords: D1 polypeptide ; Photosystem II ; psbA ; Synechocystis 6803
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transcript accumulation for the psbA, psbD, psbD-C, rbcL-S and rrn genes in Synechocystis 6803 was followed under different light conditions. psbA, psbD, psbD-C and rbcL-S transcripts required light to accumulate and the relative abundance of these transcripts differed between high and low light conditions. Under high light conditions, steady-state levels of psbA, psbD and psbD-C transcripts were higher while levels of rbcL-S transcripts were lower than under low light conditions. rrn transcripts accumulated in the dark and the transcript levels were the same under illuminated conditions. Analyses of constructed Synechocystis 6803 mutants showed that both psbA-2 and psbA-3 could produce high levels of transcripts under illuminated conditions. No psbA-1 transcripts were detected.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00016024
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  • 5
    Electronic Resource
    Electronic Resource
    Photosynthetic electron transport controls degradation but not production of psbA transcripts in the cyanobacterium Synechocystis 6803 (1991)
    Springer
    Plant molecular biology 16 (1991), S. 891-897 
    Details
    ISSN: 1573-5028
    Keywords: D1 polypeptide ; gene regulation ; psbA ; RNA stability ; Synechocystis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Accumulation and stability of psbA and rbcL-S transcripts in Synechocystis 6803 was followed in the presence and absence of the photosynthesis inhibitors DCMU and methylviologen. Our results demonstrate that both transcript production and transcript stability are important regulatory elements for psbA gene expression in Synechocystis 6803. The production of psbA transcripts was stimulated by light in a process that operated independently of the photosynthetic electron transport. However, stability of the psbA transcript increased in the dark and was controlled by photosynthetic electron transport. The psbA transcript was remarkably stable in the dark, with a half-life of approximately 7 hours. By constrast, the regulatory pattern for the rbcL-S genes was quite different. The light-stimulated production of rbcL-S transcripts was dependent on an intact photosynthetic electron transport, and rbcL-S transcript stability was higher under illuminated conditions than in darkness.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00015080
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  • 6
    Electronic Resource
    Electronic Resource
    Photosystem II characteristics of a constructedSynechocystis 6803 mutant lacking synthesis of the D1 polypeptide (1990)
    Springer
    Plant molecular biology 14 (1990), S. 1051-1054 
    Details
    ISSN: 1573-5028
    Keywords: D1 polypeptide ; psbA ; photosystem II ; polypeptide assembly ; Synechocystis 6803
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Photosystem II (PSII) composition was studied in a mutant of the cyanobacteriumSynechosystis 6803 in which synthesis of the reaction center polypeptide D1 has been inactivated. The mutant thylakoids had lost also the other reaction center polypeptide D2 and the chlorophylla-binding protein CP47. Cytochromeb559 and the chlorophylla-binding protein CP43 accumulated to almost wild-type amounts in mutant thylakoids. Also the 33 kDa polypeptide involved in water oxidation was present and membrane-bound in mutant thylakoids. The intrinsic 22 kDa polypeptide, so far known only from plants, was detected both in wild-type and mutant thylakoids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00019402
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  • 7
    Electronic Resource
    Electronic Resource
    Site-specific mutations in the D1 polypeptide affect the susceptibility of Synechocystis 6803 cells to photoinhibition (1993)
    Springer
    Plant molecular biology 22 (1993), S. 1-12 
    Details
    ISSN: 1573-5028
    Keywords: chlorophyll a fluorescence ; D1 polypeptide ; photoinhibition ; psbA genes ; site-specific mutagenesis ; Synechocystis 6803
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Photoinhibition of photosystem II in the cyanobacterium Synechocystis 6803 was followed after site-specific mutagenesis of the D1 polypeptide. Mutations were created in the stromal/cytosolic loop connecting helices D and E. Two mutations E243K and CA1, a deletion of the three glutamates 242–244 and a substitution Q241H, were made in the putative cleavage area of the D1 polypeptide. A third mutation E229D was made in the PEST-like sequence. Mutants and control cells were illuminated and FV/FM was recorded. Compared to the control, the mutants were less photoinhibited. Fluorescence relaxation after a single flash was delayed in CA1. Restoration of FV/FM after photoinhibition in the mutants was totally dependent on protein synthesis while control cells were able to recover partially also when protein synthesis was inhibited. In addition, the protein synthesis-dependent recovery of CA1 was slowed down. Our results indicate a correlation between the mutated amino acids and photoinhibition of photosystem II.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00038991
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  • 8
    Electronic Resource
    Electronic Resource
    Isolation of cDNA clones for fourteen nuclear-encoded thylakoid membrane proteins (1986)
    Springer
    Molecular genetics and genomics 204 (1986), S. 258-265 
    Details
    ISSN: 1617-4623
    Keywords: cDNA expression library ; Thylakoid membrane polypeptides ; Import in organello ; transcripts ; Spinach
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Spinach cDNA libraries, made from polyadenylated seedling RNA, have been constructed in pBR322 and the expression vector λgt11. Recombinant plasmids or phage for 14 intrinsic and peripheral thylakoid membrane proteins and one stromal protein have been identified. They encode components containing antigenic determinants against the lysine-rich 34 kd, the 23 kd and 16 kd proteins all associated with the water-splitting apparatus of the photosystem II reaction center, the ATP synthase subunits gamma, delta and CFo-II, the Rieske Fe/S protein of the cytochrome b/f complex, subunits 2, 3, 5 and 6 of the photosystem I reaction center, plastocyanin, ferredoxin oxidoreductase, chlorophyll a/b-binding apoproteins of the lightharvesting complex associated with photosystem II, and the small subunit of the stromal enzyme ribulose bisphosphate corboxylase/oxygenase. The cDNA inserts lack complementarity to plastid DNA but hybridize to restricted nuclear DNA as well as to discrete poly A+-mRNA species. The precursor products obtained after translation of hybrid selected RNA fractions in a wheat germ assay are imported and processed by isolated unbroken spinach chloroplasts. The imported components comigrate with the respective authentic proteins.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00425507
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  • 9
    Electronic Resource
    Electronic Resource
    An addressable 256×256 photodiode image sensor array with an 8-bit digital output (1993)
    Springer
    Analog integrated circuits and signal processing 4 (1993), S. 37-49 
    Details
    ISSN: 1573-1979
    Source: Springer Online Journal Archives 1860-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: Abstract We have constructed an addressable 256 × 256 photodiode sensor array together with an 8-bit ADC (analog-to-digital converter) on the same chip. Such a digital camera is easy to connect to a computer where also the flexibility of the computer can be used to control the camera output. The sensor has been constructed in two versions. The first version was implemented with a 256-column parallel-bit-slice image processor on the same die in a commercial project and the second as a separate addressable digital image sensor. The sensor was functionally fabricated using 1.6 µm design rules in a 1.2 µm CMOS process where it required a total area of 96 mm2.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01240678
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  • 10
    Electronic Resource
    Electronic Resource
    Differential expression of the psbA genes in the cyanobacterium Synechocystis 6803 (1993)
    Springer
    Molecular genetics and genomics 238 (1993), S. 161-168 
    Details
    ISSN: 1617-4623
    Keywords: D1 polypeptide ; Gene regulation ; psbA ; RNA stability ; Synechocystis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The 5′ region and transcription initiation sites of the psbA-2 and psbA-3 genes of Synechocystis 6803 were determined. The otherwise highly homologous genes were shown to diverge significantly in the 5′ noncoding regions. The transcription start site for the psbA-2 gene was mapped to position — 49 upstream of the coding region and for the psbA-3 gene to position — 88, i.e. 38 by upstream of the psbA-2 transcription start point. Both genes exhibit promoter elements, which conform in sequence and position to Escherichia coli consensus motifs. The two genes share identical — 35 sequences but differ in their — 10 sequences. Primer extension analysis demonstrated that the psbA-2 and psbA-3 genes are differentially expressed, with 〉 90 % of the total psbA transcripts being produced by the psbA-2 gene and the rest by the psbA-3 gene. Inactivation of the psbA-2 gene resulted in an eightfold up-regulation of the psbA-3 gene. The strikingly higher stability of the psbA transcripts in darkness compared to light, and the accumulation of a specific decay intermediate under dark conditions was reported previously. We show here that this dark-stability applies to both the psbA-2 and psbA-3 transcripts. The psbA-3 transcript did not appear to produce the processed intermediate, arguing for the involvement of the 5′ non-coding region as a determinant in psbA transcript degradation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00279543
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