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  • 1
    Electronic Resource
    Electronic Resource
    Parameters affecting the activity of antisense RNA sequences in tobacco protoplasts (1994)
    Springer
    Plant cell reports 13 (1994), S. 703-708 
    Details
    ISSN: 1432-203X
    Keywords: Antisense RNA ; Gene expression ; Protoplast ; Reporter gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Plasmids containing various fragments of the β-glucuronidase (GUS) gene were placed in antisense orientation downstream of the cauliflower mosaic virus 35S promoter and cotransfected with a 35S-gus construct into tobacco mesophyll protoplasts. None of the partial-length sequences were as effective as the full-length sequence in reducing GUS activity. The presence of a polyadenylation sequence downstream of the antisense sequence had an enhancing effect. The activity of the antisense sequence was largely affected by the incubation temperature of the transfected protoplasts. The chloramphenicol acetyltransferase (CAT) gene was fused to the gus coding sequence. When this construct was cotransfected with an antisense sequence directed against CAT, GUS activity was reduced. The implications of these results for the design and uses of antisense sequences are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00231628
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Characterization and expression of U1snRNA genes from potato (1992)
    Springer
    Plant molecular biology 19 (1992), S. 959-971 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; gene variants ; pre-mRNA splicing ; pseudogenes ; U1 small nuclear RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract U1 small nuclear RNAs (U1snRNAs) occur in the nucleus of plants and animals where, complexed with several proteins in the form of U1 small nuclear ribonucleoprotein particles (U1snRNPs), they play an important role in precursor messenger RNA (pre-mRNA) splicing. Ten potato U1snRNA genes have been isolated on two genomic clones illustrating the clustering of this multigene family on the potato genome. Based on both the sequence of their coding regions and upstream regulatory elements, seven of the genes are potentially functional. The other three genes were pseudogenes with defective promoter or coding region sequences. Analysis of expression of individual cloned U1snRNA genes in transfected tobacco protoplasts was impossible due to the similarity of U1snRNA sequences in tobacco. However, by marking the coding regions with oligonucleotides or constructing chimaeric genes consisting of a potato U1snRNA promoter region and maize U5snRNA coding region, three of the U1 promoter regions were shown to be transcriptionally active.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040528
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Characterization and functional analysis of Arabidopsis TFIIA reveal that the evolutionarily unconserved region of the large subunit has a transcription activation domain (1999)
    Springer
    Plant molecular biology 39 (1999), S. 515-525 
    Details
    ISSN: 1573-5028
    Keywords: TFIIA function ; transcription factor ; TBP ; Arabidopsis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract TFIIA has initially been identified as a component of transcription initiation complex of RNA polymerase II. Its role in transcription has been controversial. In this paper, we report the characterization and functional analysis of both the Arabidopsis TFIIA large and small subunits. Sequence analysis revealed that Arabidopsis TFIIA is structurally more related to animal than to yeast counterparts. Arabidopsis has at least two genes for the large subunit and one for the small subunit. Both types of genes are constitutively transcribed in various plant organs. The proteins encoded by the cDNA interact each other in yeast 2-hybrid system. Only the N-terminal part of the large subunit is necessary for the interaction with the small subunit. Recombinant Arabidopsis TFIIA polypeptides bind to TBP-DNA complex in gel shift assays. The large subunit of TFIIA can stimulate transcription in yeast and in plant cells when fused to a DNA-binding domain binding to cis sequences upstream of a minimal promoter. This trans-activating activity is localized to a 35 amino acid segment within the evolutionarily unconserved central region.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006139724849
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    Paper (German National Licenses)
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  • 4
    Electronic Resource
    Electronic Resource
    Effect of deletions in the cauliflower mosaic virus polyadenylation sequence on the choice of the polyadenylation sites in tobacco protoplasts (1991)
    Springer
    Molecular genetics and genomics 226 (1991), S. 141-144 
    Details
    ISSN: 1617-4623
    Keywords: Polyadenylation ; Gene expression ; Cauliflower mosaic virus ; Protoplasts ; Polymerase chain reaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Deletions were made in the cauliflower mosaic virus polyadenylation sequence which was cloned downstream of the β-glucuronidase gene (gus). The populations of mRNAs generated in tobacco mesophyll protoplasts by transient expression with the various constructs were analysed using a polymerase chain reaction procedure. When no deletion was present in the sequence, the mRNA appeared to be polyadenylated at two major polyadenylation sites. A deletion upstream from the AATAAA sequence made the population of polyadenylated mRNAs very heterogenous at their 3′ ends. A deletion downstream of the AATAAA sequence had no effect on the choice of the site. Alternative polyadenylation sites were used when the native polyadenylation site was deleted. These results are discussed in relation to data obtained with other polyadenylation sequences from both plants and animals.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00273597
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    Paper (German National Licenses)
    Fulltext
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