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  • 1
    Electronic Resource
    Electronic Resource
    Efficient Agrobacterium-mediated transformation of Arabidopsis thaliana using the bar gene as selectable marker (1995)
    Springer
    Plant cell reports 14 (1995), S. 450-454 
    Details
    ISSN: 1432-203X
    Keywords: Arabidopsis thaliana ; Transformation ; Phosphinothricin ; Bar ; Basta resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have established an efficient Agrobacterium-mediated transformation procedure for Arabidopsis thaliana genotype C24 using the chimeric bialaphos resistance gene (bar) coding for phosphinothricin acetyltransferase (PAT). Hypocotyl explants from young seedlings cocultivated with agrobacteria carrying a bar gene were selected on shoot-inducing media containing different concentrations of phosphinothricin (PPT) which is an active component of bialaphos. We found that 20 mg/l of PPT completely inhibited the control explants from growing whereas the explants transformed with the bar gene gave rise to multiple shoots resistant to PPT after 3 weeks under the same selection conditions. The transformation system could also be applied to root explants. Resulting plantlets could produce viable seeds in vitro within 3 months after preparation of the explants. The stable inheritance of the resistance trait, the integration and expression of the bar gene in the progeny were confirmed by genetic tests, Southern analysis and PAT enzyme assay, respectively. In addition, the mature plants in soil showed tolerance to the herbicide Basta.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00234053
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  • 2
    Electronic Resource
    Electronic Resource
    Efficient transformation of Arabidopsis thaliana: comparison of the efficiencies with various organs, plant ecotypes and Agrobacterium strains (1992)
    Springer
    Plant cell reports 12 (1992), S. 7-11 
    Details
    ISSN: 1432-203X
    Keywords: Arabidopsis thaliana ; Transformation ; Agrobacterium ; T-DNA ; Shoot regeneration ; Transgenic plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The efficiency of Agrobacterium-mediated transformation of Arabidopsis thaliana was compared with different organs, Arabidopsis ecotypes, and Agrobacterium strains. Efficiency of shoot regeneration was examined using hypocotyl, cotyledon and root explants prepared from young seedlings. Hypocotyl expiants had the highest regeneration efficiency in all of the four Arabidopsis ecotypes tested, when based on a tissue culture system of callus-inducing medium (CIM: Valvekens et al. 1988) and shoot-inducing medium (SIM: Feldmann and Marks 1986). Histochemical analysis using the ß-glucuronidase (GUS) reporter gene showed that the gusA gene expression increased as the period of preincubation on CIM was extended, suggesting that dividing cells are susceptible to Agrobacterium infection. In order to obtain transgenic shoots, hypocotyl explants preincubated for 7 or 8 days on CIM were infected with Agrobacterium containing a binary vector which carries two drug-resistant genes as selection markers, and transferred to SIM for selection of transformed shoots. Of four Arabidopsis ecotypes and of three Agrobacterium strains examined, Wassilewskija ecotype and EHA101 strain showed the highest efficiency of regeneration of transformed shoots. By combining the most efficient factors of preincubation period, Arabidopsis ecotype, tissue, and bacterial strain, we obtained a transformation efficiency of about 80–90%. Southern analysis of 124 transgenic plants showed that 44% had one copy of inserted T-DNA while the others had more than one copy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00232413
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  • 3
    Electronic Resource
    Electronic Resource
    Plant nuclear tRNAMet genes are ubiquitously interrupted by introns (1996)
    Springer
    Plant molecular biology 32 (1996), S. 427-434 
    Details
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; intron ; plant tRNAMet genes ; precursor tRNA ; splicing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated three independent clones for nuclear elongator tRNAMet genes from an Arabidopsis DNA library using a tRNAMet-specific probe generated by PCR. Each of the coding sequences for tRNAMet in these clones is identical and is interrupted by an identical 11 bp long intervening sequence at the same position in the anticodon loop of the tRNA. Their sequences differ at two positions from the intron in a soybean counterpart. Southern analysis of Arabidopsis DNA demonstrates that a gene family coding for tRNAMet is dispersed at at least eight loci in the genome. The unspliced precursor tRNAMet intermediate was detected by RNA analysis using an oligonucleotide probe complementary to the putative intron sequence. In order to know whether introns commonly interrupt plant tRNAMet genes, their coding sequences were PCR-amplified from the DNAs of eight phylogenetically separate plant species. All 53 sequences determined contain 10 to 13 bp long intervening sequences, always positioned one base downstream from the anticodon. They can all be potentially folded into the secondary structure characteristic for plant intron-containing precursor tRNAs. Surprisingly, GC residues are always present at the 5′-distal end of each intron.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00019094
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  • 4
    Electronic Resource
    Electronic Resource
    Nucleotide sequence of a putative pseudogene of histidine tRNA from Arabidopsis thaliana (1989)
    Springer
    Plant molecular biology 13 (1989), S. 721-722 
    Details
    ISSN: 1573-5028
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00016027
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  • 5
    Electronic Resource
    Electronic Resource
    Nucleotide sequence of a methionine initiator tRNA gene of Arabidopsis thaliana (1989)
    Springer
    Plant molecular biology 13 (1989), S. 599-600 
    Details
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; methionine-initiator tRNA gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027320
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  • 6
    Electronic Resource
    Electronic Resource
    Sequence analysis of three tRNAPhe nuclear genes and a mutated gene, and one gene for tRNAAla from Arabidopsis thaliana (1990)
    Springer
    Plant molecular biology 15 (1990), S. 337-346 
    Details
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; fibrinogen gene intron ; tRNAAla gene ; tRNAPhe gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three genes and one mutant gene for tRNAPhe (GAA) and one gene for tRNAAla (UGC) were isolated from a whole-cell DNA library of Arabidopsis thaliana. All three tRNAPhe genes are identical in their nucleotide sequence, but differ in their 5′ and 3′ flanking regions. The mutant tRNAPhe (GAA) gene differs from the other three genes by one nucleotide change from highly conserved G to C at the 57th nucleotide position. The primary structure of the first tRNAAla gene was also determined in this experiment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00036919
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  • 7
    Electronic Resource
    Electronic Resource
    Striking differences in mitochondrial tRNA import between different plant species (1996)
    Springer
    Molecular genetics and genomics 252 (1996), S. 404-411 
    Details
    ISSN: 1617-4623
    Keywords: Key words Mitochondria ; tRNA ; Import ; Maize ; Potato ; Larch
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A systematic comparison of the tRNAs imported into the mitochondria of larch, maize and potato reveals considerable differences among the three species. Larch mitochondria import at least eleven different tRNAs (more than half of those tested) corresponding to ten different amino acids. For five of these tRNAs [tRNAPhe(GAA), tRNALys(CUU), tRNAPro(UGG), tRNASer(GCU) and tRNASer(UGA)] this is the first report of import into mitochondria in any plant species. There are also differences in import between relatively closely related plants; wheat mitochondria, unlike maize mitochondria import tRNAHis, and sunflower mitochondria, unlike mitochondria from other angiosperms tested, import tRNASer(GCU) and tRNASer(UGA). These results suggest that the ability to import each tRNA has been acquired independently at different times during the evolution of higher plants, and that there are few apparent restrictions on which tRNAs can or cannot be imported. The implications for the mechanisms of mitochondrial tRNA import in plants are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02173005
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  • 8
    Electronic Resource
    Electronic Resource
    Molecular characterization of a gene for alanine aminotransferase from rice (Oryza sativa) (1999)
    Springer
    Plant molecular biology 39 (1999), S. 149-159 
    Details
    ISSN: 1573-5028
    Keywords: alanine aminotransferase ; gene expression ; GUS expression ; promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA clone encoding alanine aminotransferase (AlaAT) has isolated from randomly sequenced clones derived from a cDNA library of maturing rice seeds by comparison to previously identified genes. The deduced amino acid sequence was 88% and 91% homologous to those of the enzymes from barley and broomcorn millet (Panicum miliaceum), respectively. Using this cDNA as a probe, we isolated and sequenced the corresponding genomic clone. Comparison of the sequences of the cDNA and the genomic gene revealed that the coding region of the gene was interrupted by 14 introns 66 to 1547 bp long. Northern and western blotting analyses showed that the gene was expressed at high levels in developing seeds. When the 5′-flanking region between −930 and +85 from the site of initiation of transcription was fused to a reporter gene for β-glucuronidase (GUS) and then introduced into the rice genome, histochemical staining revealed strong GUS activity in the inner endosperm tissue of developing seeds and weak activity in root tips. Similar tissue-specific expression was also detected by in situ hybridization. These results suggest that AlaAT is involved in nitrogen metabolism during the maturation of rice seed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006156214716
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  • 9
    Electronic Resource
    Electronic Resource
    Splicing of Arabidopsis tRNAMet precursors in tobacco cell and wheat germ extracts (2000)
    Springer
    Plant molecular biology 44 (2000), S. 155-165 
    Details
    ISSN: 1573-5028
    Keywords: in vitro splicing ; nuclear tRNAMet introns ; suppressor tRNAMet ; tobacco cell nuclear extract ; wheat germ extract
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Intron-containing tRNA genes are exceptional within nuclear plant genomes. It appears that merely two tRNA gene families coding for tRNATyr GΨ A and elongator tRNAMet CmAU contain intervening sequences. We have previously investigated the features required by wheat germ splicing endonuclease for efficient and accurate intron excision from Arabidopsis pre-tRNATyr. Here we have studied the expression of an Arabidopsis elongator tRNAMet gene in two plant extracts of different origin. This gene was first transcribed either in HeLa or in tobacco cell nuclear extract and splicing of intron-containing tRNAMet precursors was then examined in wheat germ S23 extract and in the tobacco system. The results show that conversion of pre-tRNAMet to mature tRNA proceeds very efficiently in both plant extracts. In order to elucidate the potential role of specific nucleotides at the 3′ and 5′ splice sites and of a structured intron for pre-tRNAMet splicing in either extract, we have performed a systematic survey by mutational analyses. The results show that cytidine residues at intron-exon boundaries impair pre-tRNAMet splicing and that a highly structured intron is indispensable for pre-tRNAMet splicing. tRNA precursors with an extended anticodon stem of three to four base pairs are readily accepted as substrates by wheat and tobacco splicing endonuclease, whereas pre-tRNA molecules that can form an extended anticodon stem of only two putative base pairs are not spliced at all. An amber suppressor, generated from the intron-containing elongator tRNAMet gene, is efficiently processed and spliced in both plant extracts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006486315360
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