Nucleotide sequences of cDNA clones for B1 hordein polypeptides

Rasmussen, Søren K.; Hopp, H. Esteban; Brandt, Anders

doi:10.1007/bf02907766

Cited by 38 publications

(21 citation statements)

References 48 publications

(52 reference statements)

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“…The fourth recombinant plasmid, pcHor 2-4, contains a cDNA insert encoding one of the barley storage proteins [27]. Genes for these proteins are expressed only during seed formation and should not be transcribed in nuclei of lightgrown or etiolated leaves.…”

Section: Resultsmentioning

confidence: 99%

“…Recombinant plasmids containing sequences specific for a constitutive polypeptide of the light-harvesting chlorophyll a/b protein (pHvLF2 [l I]), the NADPH : protochlorophyllide oxidoreductase (pHvDF1 [25]), the 26-S rRNA (pc222 [26]) and B1 hordein (pcHor 2-4 [27]) all from barley, were prepared according to Birnboim and Doly [28] by two cycles ofcentrifugation in CsCl/ethidium bromide gradients.…”

Section: Preparation Of Plasmid D N Amentioning

confidence: 99%

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Phytochrome control of in vitro transcription of specific genes in isolated nuclei from barley (Hordeum vulgare)

Mösinger

Batschauer

Schäfer

et al. 1985

European Journal of Biochemistry

178

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The transcriptional rates of four different genes in shoots of barley grown under different light regimes were quantified by monitoring nuclear RNA transcripts using gene-specific hybridization probes. Isolated nuclei were pulse-labelled with [cx-~'P]UTP and the relative rates of light-harvesting chlorophyll a/b protein (LHCP) mRNA, NADPH : protochlorophyllide oxidoreductase mRNA, B1 hordein mRNA, and 26-S rRNA synthesis were measured. Irradiation of dark-grown plants with a red light pulse increased the rate of LHCP mRNA synthesis tenfold within 3 h, and the rate of rRNA synthesis more than twofold within 9 h. The relative rate of synthesis of the oxidoreductase mRNA decreased following a red light pulse reaching a minimum after 3 -6 h. As a direct proof of phytochrome involvement in the light-induced stimulation of LHCP and the repression of the oxidoreductase transcripts for both responses, red/far-red reversibility could be demonstrated. We conclude that phytochrome is able both to increase the transcription of certain nuclear genes and decrease the transcription of others.The light-dependent control of plant development is one of the central themes of current plant biology [I]. Light has been shown to induce an increase in a number of mRNA species encoding, for instance, several plastid-specific proteins like the light-harvesting chlorophyll a/b protein and the small subunit of the ribulose-I ,5-bisphosphate carboxylase [2-131. At the same time the concentration of other transcripts, e.g. those coding for the NADPH : protochlorophyllide oxidoreductase (reductase) and phytochrome, is drastically reduced upon illumination of darkgrown plants [14-171. In both instances the reciprocal effect of light on the level of these two groups of mRNAs is mediated by the same photoreceptor phytochrome [3, 5, 6, 8 -1 1,13 -171. These light effects could occur by two possible mechanisms : a phytochrome-controlled change in the rate of transcription initiation and/or a phytochrome-induced change in the stability of transcripts. In the present study we have concentrated on elucidating the first of these two mechanisms. Isolated nuclei of barley have been pulse-labelled and the newly synthesized transcripts analyzed. Available evidence suggests that the majority of RNA synthesis in isolated nuclei represents elongation of already initiated RNA chains and that the rates of transcription in pulse-labelled isolated nuclei can be monitored accurately without the complications of RNA processing or stabilization which occurs within the intact cell [18]. In previous studies it could be demonstrated [I91 that phytochrome controls the overall transcription rate in isolated Avena nuclei. Studies on the synthesis of specific transcripts in isolated nuclei of pea and duckweed, a lightinduced increase in the rate of transcription of genes encoding the light-harvesting chlorophyll a/b protein and the small subunit of the ribulose-l,5-bisphosphate carboxylase has been described [12, 131. This light effect was mediated by phytochrome [13]. As...

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Section: Resultsmentioning

confidence: 99%

Section: Preparation Of Plasmid D N Amentioning

confidence: 99%

Phytochrome control of in vitro transcription of specific genes in isolated nuclei from barley (Hordeum vulgare)

Mösinger

Batschauer

Schäfer

et al. 1985

European Journal of Biochemistry

178

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“…In the two known B-hordein polypeptides this region is localized in the NHz-terminal third of the molecule. The remaining part appears to be unique to the B-hordein polypeptide and amino acid substitutions in this region appear to be the major source for the charge polymorphism observed among the B-hordein polypeptides (5,36). In contrast, the glutamine + proline rich region occupies most of the C-hordein polypeptides while only short regions in the NH2-and COOH-terminal part are unique.…”

Section: Introductionmentioning

confidence: 98%

Monoclonal antibodies to hordein polypeptides

Ullrich

Rasmussen

Høyer‐Hansen

et al. 1986

Carlsberg Res. Commun.

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Monoclonal antibodies to hordein polypeptides have been produced and characterized. All antibodies reacted with more than one polypeptide, indicating that different hordein polypeptides have common epitopes. Of the seventeen isolated hybridoma lines, six secreted antibodies which reacted both with B-and C-hordein, five recognized only B-hordein and one secreted antibodies specific for C-hordein polypeptides. Two clones produced antibodies to an unidentified polypeptide doublet with an apparent molecular weight of 43 kD. These two polypeptides have at least one epitope in common with B-hordein polypeptides and are not extractable in water but in alcohol. They are not visible by Coomassie blue staining.The mutant M56 fails to synthesize B-hordein polypeptides due to a major deletion. It contains polypeptides in the B-hordein region of the gel, which have epitopes recognized by the B-hordein antibodies, but are different from B-hordeins. The 43 kD hordein doublet was more abundant in the mutant than in the wild-type endosperm, suggesting that these polypeptides like the C-hordein polypeptides are synthesized in larger amounts as a consequence of the absent B-hordein polypeptide synthesis. Mutant 1508 which has an impaired synthesis of both B-and C-hordein polypeptides was also found to be defective in the synthesis of the 43 kD polypeptide doublet. It was shown that immunofluoreseent techniques may be of practical importance to identify mutants with elevated or reduced ~tmounts of specific hordein polypeptides. Due to the higher sensitivity compared to SDS-PAGE, immunoblotting can be a useful tool for genotype identification in breeding programmes.

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“…This was verified by Anders Brandt, John Ingversen, Verena Cameron-Mills, and Susan Madrid using isolated endoplasmatic reticulum (5,6,(11)(12)(13) and isolated protein bodies and by localization studies with monoclonal antibodies developed by Gunilla Høyer-Hansen (58,65,66,77) (see Supplemental Figures 2 and 3). Cloning and sequencing of structural genes for B-, C-, and γ-hordein polypeptides have corroborated that the Hor 1, Hor 2, and Hor F loci each consist of a family of closely linked structural genes for these proteins, with each gene having its own promoter (7,10,18,24,62,63,70). Some of these genes contain stop codons, but transient transformation experiments revealed that expression of such a gene in the endosperm can take place by amber codon suppression (19).…”

Section: Prefacementioning

confidence: 99%

From Analysis of Mutants to Genetic Engineering

Wettstein

2007

Annu. Rev. Plant Biol.

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This chapter describes the research of developing transgenic barley for synthesis of recombinant proteins with practical significance and of metabolic engineering of proanthocyanidin-free barley. The results were obtained by graduate students, postdoctoral researchers, and visiting scientists at the Carlsberg Laboratory from 1972-1996 and during the past ten years at Washington State University. It is written in appreciation of their enthusiasm, skill, and perseverance.

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Nucleotide sequences of cDNA clones for B1 hordein polypeptides

Cited by 38 publications

References 48 publications

Phytochrome control of in vitro transcription of specific genes in isolated nuclei from barley (Hordeum vulgare)

Phytochrome control of in vitro transcription of specific genes in isolated nuclei from barley (Hordeum vulgare)

Monoclonal antibodies to hordein polypeptides

From Analysis of Mutants to Genetic Engineering

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