Vectors with rare-cutter restriction enzyme sites for expression of open reading frames in transgenic plants

Cloning of long open reading frames (ORFs) into plant gene expression vectors and transfer of the chimeric expression cassettes into binary vectors is often hampered by the presence of restriction enzyme cleavage sites internal to the open reading frame (ORF) to be expressed. We therefore modified the commonly used expression vector pRT100 [7] and several pGPTV binary vectors [2] by replacing 6 bp restriction sites with 8 bp sequences recognized by rare-cutter restriction enzymes.