Early and multiple Ac transpositions in rice suitable for efficient insertional mutagenesis View Full Text


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Article Info

DATE

2001-05

AUTHORS

Raffaella Greco, Pieter B.F. Ouwerkerk, Anke J.C. Taal, Cristina Favalli, Thierry Beguiristain, Pere Puigdomènech, Lucia Colombo, J. Harry C. Hoge, Andy Pereira

ABSTRACT

A GFP excision assay was developed to monitor the excision of Ac introduced into rice by Agrobacterium-mediated transformation. The presence of a strong double enhancer element of the CaMV 35S promoter adjacent to the Ac promoter induced very early excision, directly after transformation into the plant cell, exemplified by the absence of Ac in the T-DNA loci. Excision fingerprint analysis and characterization of transposition events from related regenerants revealed an inverse correlation between the number of excision events and transposed Ac copies, with single early excisions after transformation generating Ac amplification. New transpositions were generated at a frequency of 15–50% in different lines, yielding genotypes bearing multiple insertions, many of which were inherited in the progeny. The sequence of DNA flanking Ac in three representative lines provided a database of insertion tagged sites suitable for the identification of mutants of sequenced genes that can be examined for phenotypes in a reverse genetics strategy to elucidate gene function. Remarkably, two-thirds of Ac tagged sites showing homology to sequences in public databases were in predicted genes. A clear preference of transposon insertions in genes that are either predicted by protein coding capacity or by similarity to ESTs suggests that the efficiency of recovering knockout mutants of genes could be about three times higher than random. Linked Ac transposition, suitable for targeted tagging, was documented by segregation analysis of a crippled Ac element and by recovery of a set of six insertions in a contiguous sequence of 70 kb from chromosome 6 of rice. More... »

PAGES

215-227

References to SciGraph publications

  • 1997-09. Transposon tagging in rice in PLANT MOLECULAR BIOLOGY
  • 1991-03. The use of transgenic plants to understand transposition mechanisms and to develop transposon tagging strategies in PLANT MOLECULAR BIOLOGY
  • 1999-12. Sequence and analysis of chromosome 2 of the plant Arabidopsis thaliana in NATURE
  • 1999-12. Sequence and analysis of chromosome 4 of the plant Arabidopsis thaliana in NATURE
  • 2000-12. Analysis of the genome sequence of the flowering plant Arabidopsis thaliana in NATURE
  • 1993-06. Trans-activation and stable integration of the maize transposable element Ds cotransfected with the Ac transposase gene in transgenic rice plants in MOLECULAR GENETICS AND GENOMICS
  • 1994-03. The presence of enhancers adjacent to the Ac promoter increases the abundance of transposase mRNA and alters the timing of Ds excision in Arabidopsis in PLANT MOLECULAR BIOLOGY
  • 1998-03. Soluble, highly fluorescent variants of green fluorescent protein (GFP) for use in higher plants in PLANT MOLECULAR BIOLOGY
  • 1989-05. Induced transposition of Ds by a stable Ac in crosses of transgenic tobacco plants in MOLECULAR GENETICS AND GENOMICS
  • 1990-03-01. Production of Correctly Processed Human Serum Albumin in Transgenic Plants in BIO/TECHNOLOGY
  • 1995-09. A two-element Enhancer-Inhibitor transposon system in Arabidopsis thaliana in MOLECULAR GENETICS AND GENOMICS
  • 1991-07. Introduction and transposition of the maize transposable element Ac in rice (Oryza sativa L.) in MOLECULAR GENETICS AND GENOMICS
  • 1994-02. Efficient plant regeneration through somatic embryogenesis from callus induced on mature rice embryos (Oryza sativa L.) in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1023/a:1010607318694

    DOI

    http://dx.doi.org/10.1023/a:1010607318694

    DIMENSIONS

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/11442061


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    41 Agrobacterium
    42 CaMV 35S promoter
    43 DNA
    44 DNA loci
    45 ESTs
    46 Excision fingerprint analysis
    47 GFP excision
    48 absence
    49 absence of AC
    50 amplification
    51 analysis
    52 capacity
    53 cells
    54 characterization
    55 chromosome 6
    56 clear preference
    57 coding capacity
    58 contiguous sequence
    59 copies
    60 correlation
    61 crippled Ac element
    62 database
    63 database of insertion
    64 different lines
    65 double enhancer element
    66 early excision
    67 efficiency
    68 efficient insertional mutagenesis
    69 elements
    70 enhancer elements
    71 events
    72 excision
    73 excision events
    74 excision of Ac
    75 fingerprint analysis
    76 frequency
    77 function
    78 gene function
    79 genes
    80 genetic strategies
    81 genotypes
    82 homology
    83 identification
    84 identification of mutants
    85 insertion
    86 insertional mutagenesis
    87 inverse correlation
    88 kb
    89 knockout mutants
    90 lines
    91 loci
    92 multiple Ac transpositions
    93 multiple insertions
    94 mutagenesis
    95 mutants
    96 new transpositions
    97 number
    98 phenotype
    99 plant cells
    100 preferences
    101 presence
    102 progeny
    103 promoter
    104 protein coding capacity
    105 public databases
    106 recovery
    107 regenerants
    108 related regenerants
    109 representative lines
    110 reverse genetic strategy
    111 rice
    112 segregation analysis
    113 sequence
    114 sequence of DNA
    115 sequenced gene
    116 set
    117 similarity
    118 single early excisions
    119 sites
    120 strategies
    121 strong double enhancer element
    122 tagged sites
    123 tagging
    124 targeted tagging
    125 time
    126 transformation
    127 transposition
    128 transposition events
    129 transposon insertion
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