PROTEIN EXPRESSION IN PLANTS

摘要

The present invention relates to protein expression in plants, particularly the large-scale production of recombinant polypeptides in whole Nicotiana tabacum plants. The use of preselected combination of N. tabacum varieties and Agrobacterium strains, optionally including one or more improvements to the transient expression-based methods of the invention, enables the production of large quantities of a heterologous polypeptides economically and in a short period of time.

主权项

1. A method for producing a heterologous polypeptide in Nicotiana tabacum comprising the steps of:
(i) providing a combination of a selected variety, breeding line, or cultivar of a Nicotiana tabacum plant and a selected strain of an Agrobacterium species, which variety, breeding line, or cultivar, exhibits less than 10% necrosis 5 days after leaves of said variety, breeding line, or cultivar have been injected by syringe with the selected Agrobacterium strain at a cell density of OD600 of 0.32; (ii) infiltrating a whole plant of the selected variety, breeding line, or cultivar of Nicotiana tabacum with a suspension of the selected strain of the Agrobacterium species at an OD600 of between 0.1 and 4.0, said strain comprising an expressible nucleotide sequence encoding the polypeptide; and (iii) incubating the infiltrated plant for a period of between 5 days and 10 days under conditions that allow expression of the expressible nucleotide sequence in the infiltrated plant and accumulation of the heterologous polypeptide.