Expression of biologically active human granulocyte macrophage colony stimulating factor in the seeds of transgenic tobacco.

Abstract

The feasibility of producing recombinant (rt) and biologically active granulocyte-macrophage colony stimulating factor (GM-CSF) in the seeds of transgenic tobacco plants was investigated. The rice seed-specific glutelin promoter (Gt1) was used to direct the expression of the human (h) GM-CSF coding sequence in tobacco seeds. Transgenic tobacco plants producing rthGM-CSF were compared in biological assays with tobacco plants expressing a glutelin/rthGM-CSF fusion protein, driven by the Gt3 promoter. The T7 Sequencing kit from Pharmacia was used to sequence and confirm the authenticity of the Gt1 expression construct. The glutelin-1 signal sequence was fused in the correct orientation to the hGM-CSF cDNA. The rthGM-CSF expression cassette (2.5 kb) was subcloned in a plant binary vector pRD400, which contained a kanamycin resistance gene. The pRD400 vector containing the 2.5 kb construct was used to transform Agrobacterium tumefaciens cells. Tobacco (Nicotiana tabacum cv. Xanthi) leaf sections were transformed by A. tumefaciens carrying the complete 2.5 kb construct. (Abstract shortened by UMI.)