The plant exon finder: A tool for precise detection of exons using a T-DNA-based tagging approach

Fumito Jingu, Toshiyuki Shirase, Ichiro Ohtomo, Akihiro Imai, Yoshibumi Komeda, Taku Takahashi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

We have developed a new tool, named the plant exon finder (PEF), for identifying exons in plant genome sequences as an applied technique of T-DNA insertional mutagenesis. The T-DNA constructs contain a heat-shock gene promoter or the cauliflower mosaic virus (CaMV) 35S promoter, followed by the first exon of an Arabidopsis gene with its start codon and the intron donor sequence facing the T-DNA left border (LB) in order to trap exons in the genome. The constructs were used to make transgenic Arabidopsis plants. We generated 280 transgenic lines and identified 156 T-DNA-tagged readthrough transcripts by reverse transcriptase-polymerase chain reaction (RT-PCR) using an oligo(dT)-linker primer and a T-DNA-specific primer. Sequence analysis of the RT-PCR products revealed that 18 of them carried cDNAs processed by the use of an intron acceptor sequence adjacent to T-DNA insertion sites and 11 of them were in-frame fusions. In one case, the readthrough transcript trapped an exon located 1.6 kb downstream of the site of the insertion.

Original languageEnglish
Pages (from-to)267-273
Number of pages7
JournalGene
Volume338
Issue number2
DOIs
Publication statusPublished - Sep 1 2004
Externally publishedYes

Fingerprint

Exons
Reverse Transcriptase Polymerase Chain Reaction
Arabidopsis
Introns
Caulimovirus
Plant Genome
DNA Primers
Initiator Codon
Insertional Mutagenesis
Genetically Modified Plants
Genes
Sequence Analysis
T-DNA
Shock
Complementary DNA
Hot Temperature
Genome

Keywords

  • Arabidopsis
  • BAC
  • bacterial artificial chromosome
  • Gene trap
  • GFP
  • green fluorescent protein
  • heat shock
  • Heat-shock promoter
  • HS
  • LB
  • left border
  • PEF
  • plant exon finder
  • RT-PCR

ASJC Scopus subject areas

  • Genetics

Cite this

The plant exon finder : A tool for precise detection of exons using a T-DNA-based tagging approach. / Jingu, Fumito; Shirase, Toshiyuki; Ohtomo, Ichiro; Imai, Akihiro; Komeda, Yoshibumi; Takahashi, Taku.

In: Gene, Vol. 338, No. 2, 01.09.2004, p. 267-273.

Research output: Contribution to journalArticle

Jingu, Fumito ; Shirase, Toshiyuki ; Ohtomo, Ichiro ; Imai, Akihiro ; Komeda, Yoshibumi ; Takahashi, Taku. / The plant exon finder : A tool for precise detection of exons using a T-DNA-based tagging approach. In: Gene. 2004 ; Vol. 338, No. 2. pp. 267-273.
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