title: High levels of expression of fibroblast growth factor 21 in transgenic tobacco (Nicotiana benthamiana).
abstract: Fibroblast growth factor-21 (FGF21) is a hepatic hormone that plays a critical role in metabolism, stimulating fatty acid oxidation in the liver and glucose uptake in adipose tissue. In this study, we produced tobacco plants expressing human recombinant FGF21 (hFGF21) via Agrobacterium-mediated transformation using a potato virus X (PVX)-based vector (pgR107). The vector contained the sequence encoding the human FGF21 gene fused with green florescence protein and a histidine tag. The recombinant plasmid was introduced into leaf cells of Nicotiana benthamiana (a wild Australian tobacco) via Agrobacterium-mediated agroinfiltration. As determined by fluorescence and Western blot of leaf extracts, the hFGF21 gene was correctly translated in tobacco plants. Seven days after agroinfection, the recombinant hFGF21 had accumulated to levels as high as 450 μg g(-1) fresh weight in leaves of agroinfected plants. The recombinant hFGF21 was purified from plant tissues by Ni-NTA affinity chromatography, and the purified hFGF21 stimulated glucose uptake in 3T3/L1 cells. This indicated that the recombinant hFGF21 expressed via the PVX viral vector in N. benthamiana was biologically active.
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin 130118, China.
authors.forename: Hongqi
authors.initials: H
authors.lastname: Fu
authors.forename: Shifeng
authors.initials: S
authors.lastname: Pang
authors.forename: Ping
authors.initials: P
authors.lastname: Xue
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.forename: Xiuming
authors.initials: X
authors.lastname: Liu
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Wang
authors.forename: Tingting
authors.initials: T
authors.lastname: Li
authors.forename: Haiyan
authors.initials: H
authors.lastname: Li
authors.forename: Xiaokun
authors.initials: X
authors.lastname: Li
mesh_terms: D041721:3T3-L1 Cells; D016960:Agrobacterium tumefaciens / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005346:Fibroblast Growth Factors / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry*; D005947:Glucose / Q000378:metabolism; D006801:Humans; D051379:Mice; D018515:Plant Leaves / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D010957:Plasmids; D017863:Potexvirus / Q000235:genetics* / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism / Q000821:virology
chemical_list: D011993:Recombinant Fusion Proteins; C414620:fibroblast growth factor 21; D005346:Fibroblast Growth Factors; D005947:Glucose