Fluorescence in situ hybridization of single copy transgenes in rice chromosomes Academic Article uri icon

abstract

  • Fluorescence in situ hybridization (FISH) is a powerful tool for visualizing the chromosomal location of targeted sequences and has been applied in many areas, including karyotyping, breeding and characterization of genes introduced into the plant genome. A simple, routine and sensitive FISH procedure was developed for localizing single copy genes in rice (Oryza sativa L.) metaphase chromosomes. We used digoxygenin-labeled endogenous or T-DNA sequences as small as 5.6 kb to probe corresponding endogenous sequences or the T-DNA insert in denatured rice metaphase chromosomes prepared from root meristem tissue. The hybridized probe sequence was labeled with cy3-conjugated anti-mouse IgG and visualized using fluorescence microscopy. Single copy and multiple copy introduced T-DNA sequences, as well as endogenous sequences, were localized on the chromosomes. The FISH protocol was effectively used to screen the chromosomal location of introduced T-DNA and number of integration loci in rice.

author list (cited authors)

  • Kharb, P., Dong, J. J., Islam-Faridi, M. N., Stelly, D. M., & Hall, T. C.

citation count

  • 8

publication date

  • January 2001

keywords

  • Digoxygenin (dig)
  • Fluorescence In Situ Hybridization (fish)
  • Oryza Sativa L
  • T-dna
  • Transgenic