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  • 1
    Electronic Resource
    Electronic Resource
    Isolation and molecular characterization of dTnp1, a mobile and defective transposable element of Nicotiana plumbaginifolia (1994)
    Springer
    Molecular genetics and genomics 242 (1994), S. 194-200 
    Details
    ISSN: 1617-4623
    Keywords: Transposable element ; Nitrate reductase ; Nicotiana plumbaginifolia ; γ-Ray mutagenesis ; Nucleotide sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By Northern blot analysis of nitrate reductase-deficient mutants of Nicotiana plumbaginifolia, we identified a mutant (mutant D65), obtained after γ-ray irradiation of protoplasts, which contained an insertion sequence in the nitrate reductase (NR) mRNA. This insertion sequence was localized by polymerase chain reaction (PCR) in the first exon of NR and was also shown to be present in the NR gene. The mutant gene contained a 565 by insertion sequence that exhibits the sequence characteristics of a transposable element, which was thus named dTnp1. The dTnp1 element has 14 by terminal inverted repeats and is flanked by an 8-bp target site duplication generated upon transposition. These inverted repeats have significant sequence homology with those of other transposable elements. Judging by its size and the absence of a long open reading frame, dTnp1 appears to represent a defective, although mobile, transposable element. The octamer motif TTTAGGCC was found several times in direct orientation near the 5′ and 3′ ends of dTnp1 together with a perfect palindrome located after the 5′ inverted repeat. Southern blot analysis using an internal probe of dTnp1 suggested that this element occurs as a single copy in the genome of N. plumbaginifolia. It is also present in N. tabacum, but absent in tomato or petunia. The dTnp1 element is therefore of potential use for gene tagging in Nicotiana species.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00391013
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  • 2
    Electronic Resource
    Electronic Resource
    Plant cyclins: a unified nomenclature for plant A-, B- and D-type cyclins based on sequence organization (1996)
    Springer
    Plant molecular biology 32 (1996), S. 1003-1018 
    Details
    ISSN: 1573-5028
    Keywords: cyclins ; nomenclature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The comparative analysis of a large number of plant cyclins of the A/B family has recently revealed that plants possess two distinct B-type groups and three distinct A-type groups of cyclins [1]. Despite earlier uncertainties, this large-scale comparative analysis has allowed an unequivocal definition of plant cyclins into either A or B classes. We present here the most important results obtained in this study, and extend them to the case of plant D-type cyclins, in which three groups are identified. For each of the plant cyclin groups, consensus sequences have been established and a new, rational, plant-wide naming system is proposed in accordance with the guidelines of the Commission on Plant Gene Nomenclature. This nomenclature is based on the animal system indicating cyclin classes by an upper-case roman letter, and distinct groups within these classes by an arabic numeral suffix. The naming of plant cyclin classes is chosen to indicate homology to their closest animal class. The revised nomenclature of all described plant cyclins is presented, with their classification into groups CycA1, CycA2, CycA3, CycB1, CycB2, CycD1, CycD2 and CycD3.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00041384
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  • 3
    Electronic Resource
    Electronic Resource
    Molecular cloning and characterisation of the two homeologous genes coding for nitrate reductase in tobacco (1989)
    Springer
    Molecular genetics and genomics 216 (1989), S. 10-15 
    Details
    ISSN: 1617-4623
    Keywords: Nicotiana tabacum ; Nitrate reductase ; Structural gene ; Transcription ; Sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The two structural genes encoding tobacco nitrate reductases (NR) were isolated from tobacco genomic libraries constructed in lambda EMBL phages. Two independent genomic clones of 12.6 and 13.5 kbp, respectively, cross-hybridizing with a partial tobacco NR cDNA probe, were further characterized. Southern blot experiments were performed with the NR cDNA probe on genomic DNA derived from Nicotiana tabacum and from the ancestors of tobacco, N. sylvestris and N. tomentosiformis. They showed that the larger clone, referred to as nia-1, was related to the N. tomentosiformis parent, and the smaller one, referred to as nia-2, to the N. sylvestris parent. Both homeologous genes were found to be expressed in tobacco. The sequence of the gene nia-2, from which the cDNA previously cloned is derived, was determined. It encodes a 904 amino acid protein. Three intervening sequences were found interspersed with the coding sequence of the enzyme. The precise location of the transcription initiation site on the structural gene was mapped by primer extension experiments. A TATA consensus sequence was detected 32 bp upstream from the transcription initiation site. The leader sequence of the transcript is 138 nucleotides long and a stable secondary structure involving the translation initiation site has been proposed. The amino acid sequence of tobacco NR deduced from the nucleotide sequence of the gene shows that heme and FAD binding domains occupy the entire C-terminal moiety of the polypeptide. The remaining N-terminal part of the protein should thus carry the catalytic site of nitrate reduction by the molybdenum cofactor.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00332224
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  • 4
    Electronic Resource
    Electronic Resource
    Cloning of DNA fragments complementary to tobacco nitrate reductase mRNA and encoding epitopes common to the nitrate reductases from higher plants (1987)
    Springer
    Molecular genetics and genomics 209 (1987), S. 552-562 
    Details
    ISSN: 1617-4623
    Keywords: Nitrate reductase ; cDNA expression cloning ; Tobacco ; Sequence ; Cytochrome b5
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Messenger RNAs encoding the nitrate reductase apoenzyme from tobacco can be translated in a cell-free system. Poly(A)+ mRNA fractions from the 23-32 S area of a sucrose gradient were used to build a cDNA library in the expression vector λgt11 with an efficiency of cloning of approximately 104 recombinants/ng mRNA. Recombinant clones were screened with a rabbit polyclonal antibody directed against the corn nitrate reductase, which cross reacts specifically with the nitrate reductases from dicotyledons. Among 240000 recombinant plaques, eight clones were isolated containing inserts of sizes ranging from 1.6 kb to 2.1 kb and sharing sequence homologies. Seven of these clones contained a common internal 1.6 kb EcoRI fragment. The identity of these clones was confirmed as follows. A fusion protein of 170 kDa inducible by IPTG and recognized by the rabbit nitrate reductase antibody was expressed by a lysogen derived from one of the recombinants. The antibodies binding the fused protein were eluted and shown to be inhibitory to the catalytic activity of tobacco nitrate reductase. Two monoclonal antibodies directed against nitrate reductase were also able to bind the hybrid protein. The 1.6 kb EcoRI fragment was sequenced by the method of Sanger. The open reading frame corresponding to a translational fusion with the β-galactosidase coding sequence of the vector shared strong homology at the amino acid level with the heme-binding domain of proteins of the cytochrome b5 superfamily and with human erythrocyte cytochrome b5 reductase. When the 1.6 kb EcoRI fragment was used as a probe for Northern blot experiments a signal corresponding to a 3.5 kb RNA was detected in tobacco and in Nicotiana plumbaginifolia mRNA preparations but no cross-hybridization with corn mRNAs was detected. The probe hybridized with low copy number sequences in genomic blots of tobacco DNA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00331162
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  • 5
    Electronic Resource
    Electronic Resource
    Analysis of T-DNA-mediated translational β-glucuronidase gene fusions (1998)
    Springer
    Plant molecular biology 36 (1998), S. 205-217 
    Details
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; Arabidopsis thaliana ; gene fusion ; insertion mutagenesis ; hybrid proteins ; serine/threonine receptor kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three random translational β-glucuronidase (gus) gene fusions were previously obtained in Arabidopsis thaliana, using Agrobacterium-mediated transfer of a gus coding sequence without promoter and ATG initiation site. These were analysed by IPCR amplification of the sequence upstream of gus and nucleotide sequence analysis. In one instance, the gus sequence was fused, in inverse orientation, to the nos promoter sequence of a truncated tandem T-DNA copy and translated from a spurious ATG in this sequence. In the second transgenic line, the gus gene was fused to A. thaliana DNA, 27 bp downstream an ATG. In this line, a large deletion occurred at the target site of the T-DNA. In the third line, gus is fused in frame to a plant DNA sequence after the eighth codon of an open reading frame encoding a protein of 619 amino acids. This protein has significant homology with animal and plant (receptor) serine/threonine protein kinases. The twelve subdomains essential for kinase activity are conserved. The presence of a potential signal peptide and a membrane-spanning domain suggests that it may be a receptor kinase. These data confirm that plant genes can be tagged as functional translational gene fusions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1005902730810
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