The utility of flow sorting to identify chromosomes carrying a single copy transgene in wheat.

ABSTRACT:

Background

Identification of transgene insertion sites in plant genomes has practical implications for crop breeding and is a stepping stone to analyze transgene function. However, single copy sequences are not always easy to localize in large plant genomes by standard approaches.

Results

We employed flow cytometric chromosome sorting to determine chromosomal location of barley sucrose transporter construct in three transgenic lines of common wheat. Flow-sorted chromosomes were used as template for PCR and fluorescence in situ hybridization to identify chromosomes with transgenes. The chromosomes carrying the transgenes were then confirmed by PCR using DNA amplified from single flow-sorted chromosomes as template.

Conclusions

Insertion sites of the transgene were unam

SUBMITTER: Capal P 

PROVIDER: S-EPMC4845436 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature