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Isolation and molecular characterization of dTnp1, a mobile and defective transposable element of Nicotiana plumbaginifolia

Molecular and General Genetics MGG volume 242, pages 194–200 (1994)Cite this article

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.

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Affiliations

  1. Laboratoire de Biologie Cellulaire, INRA, 78026, Versailles Cedex, France

    Christian Meyer, Sylvie Pouteau, Pierre Rouzé & Michel Caboche

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  1. Christian Meyer
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  2. Sylvie Pouteau
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  3. Pierre Rouzé
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  4. Michel Caboche
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Communicated by H. Böhme

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Meyer, C., Pouteau, S., Rouzé, P. et al. Isolation and molecular characterization of dTnp1, a mobile and defective transposable element of Nicotiana plumbaginifolia . Molec. Gen. Genet. 242, 194–200 (1994). https://doi.org/10.1007/BF00391013

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  • DOI: https://doi.org/10.1007/BF00391013

Key words

  • Transposable element
  • Nitrate reductase
  • Nicotiana plumbaginifolia
  • γ-Ray mutagenesis
  • Nucleotide sequence