[A quick method to estimate the T-DNA copy number in transgenic rice using inverse PCR (IPCR)]

Shi Yan Sheng Wu Xue Bao. 1999 Jun;32(2):207-11.
[Article in Chinese]

Abstract

In order to obtain single T-DNA copy transgenic rice, we have established a quick method to estimate the T-DNA copy number in transgenic rice using inverse PCR (IPCR). IPCR was used to amplify junction fragments, i.e. plant genomic DNA sequences flanking the known T-DNA sequences, which will help to estimate the T-DNA copy number in transgenic rice. We have analyzed 20 transgenic plants of 15 transgenic lines. Most plants (12) contain one integrated T-DNA copy per genome, 3 plants contain two and 1 plant contains 3 copies. In 4 transgenic plants no T-DNA copies could be detected using this method. The IPCR results were further tested by Southern analysis and sequence analysis.

Publication types

  • English Abstract

MeSH terms

  • DNA, Bacterial / analysis*
  • Gene Dosage
  • Oryza / genetics*
  • Plants, Genetically Modified
  • Polymerase Chain Reaction / methods*

Substances

  • DNA, Bacterial
  • T-DNA