A convenient set of vectors for expression of multiple gene combinations in plants.

摘要

A set of vectors has been developed to simplify shuttling expression cassettes between small plasmids of high copy number. The vectors are ideal for experiments involving biolistic transient expression and a binary transformation plasmid. Three cassettes for the expression of a cloned coding sequence behind different promoters have been modified; combinations of these cassettes can be excised with NotI, and sequentially cloned into the transformation vector in a procedure that removes the first cloning site. The system was demonstrated by inducing anthocyanin synthesis with a paired set of regulatory genes of maize via particle bombardment of a maize cell suspension. Both chimaeric genes were then excised and sequentially assembled into a binary vector to create the cassette used to successfully transform tobacco plants via Agrobacterium-mediated transformation. The system has general applicability in facilitating biolistic experiments during the initial construction phase, and in simplifying mixing and matching of different promoters and coding sequences in the transgenic phase.