[Amplification and analysis of T-DNA flanking sequences in transgenic rice]

Yi Chuan Xue Bao. 2001;28(4):345-51.
[Article in Chinese]

Abstract

Rice transformation mediated by Agrobacterium tumefaciens has technically matured to some extent, but the mechanics of T-DNA integration in transgenic rice remains largely unknown. Using thermal asymmetric interlaced PCR (TAIL-PCR), we analyzed the flanking sequences of T-DNAs in transgenic rice plants, in which the resistance gene for rice bacterial blight disease, Xa21, had been integrated stably. Sequence analysis of 24 fragments amplified by TAIL-PCR showed that of them 14 were rice genomic DNA, 9 contained vector backbone sequences, and one was a fragment of the exogenous gene Xa21. The characteristics of the 14 rice genomic DNA sequences at T-DNA integrated sites are significantly different from those of the reported rice genomic sequences into which exogenous genes were integrated through direct DNA transformation methods. T-DNA border sequences integrated in rice genome have similar features as those integrated in dicotyledonous genomes. In the backbone containing flanking sequences (37.5%, 9/24) vector backbones appeared in different types.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA, Bacterial / chemistry*
  • Molecular Sequence Data
  • Oryza / genetics*
  • Plants, Genetically Modified
  • Polymerase Chain Reaction
  • Transformation, Genetic

Substances

  • DNA, Bacterial
  • T-DNA