High-level expression of basic fibroblast growth factor in transgenic soybean seeds and characterization of its biological activity.
The glycinin G1 gene encodes a soybean seed storage protein accumulating at a high level. We have used the G1 promoter to confer seed-specific expression of human basic fibroblast growth factor (bFGF) in transgenic soybeans. The coding region of 18 kDa bFGF was fused to the promoter or promoter-signal peptide sequence of G1 gene, and transferred into soybean. Analysis of transgenic plants demonstrated that bFGF transcript or protein was confined to the seeds. The highest level of bFGF accumulation in the seeds reached up to 2.3% of total soluble protein. The soybean-derived bFGF was biologically active as confirmed by its mitogenic activity on Balb/c 3T3 cells, and exhibited other properties identical to native bFGF. We also observed a seed-specific expression of beta-glucuronidase driven by the G1 promoter. These results indicated that the G1 promoter contains essential cis-elements for seed-specific expression, and thus can be used for expression of pharmaceutical proteins in soybean seeds.
Ding
Shi-Hua
SH
Lab of Biotechnology, Collage of Fishery Science, Southwest University, Chongqing, China.
Huang
Li-Ying
LY
Wang
Yi-Ding
YD
Sun
Han-Chang
HC
Xiang
Zhong-Huai
ZH
D000818:Animals; D041702:BALB 3T3 Cells; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D005916:Globulins / Q000235:genetics; D006801:Humans; D051379:Mice; D008934:Mitogens / Q000096:biosynthesis*; D030821:Plants, Genetically Modified / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012639:Seeds / Q000235:genetics / Q000378:metabolism*; D030262:Soybean Proteins / Q000235:genetics; D013025:Glycine max / Q000235:genetics*
D005916:Globulins; D008934:Mitogens; D030262:Soybean Proteins; D016222:Fibroblast Growth Factor 2; C011474:glycinin