Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin 130118, China.
Appl Biochem Biotechnol. 2011 Sep;165(2):465-75. doi: 10.1007/s12010-011-9265-4. Epub 2011 Apr 20.
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.
成纤维细胞生长因子 21（FGF21）是一种肝脏激素，在代谢中起着关键作用，刺激肝脏中的脂肪酸氧化和脂肪组织中的葡萄糖摄取。在这项研究中，我们通过使用基于马铃薯病毒 X（PVX）的载体（pgR107），通过农杆菌介导的转化，在烟草植物中表达了人重组 FGF21（hFGF21）。该载体包含与人 FGF21 基因融合的绿色荧光蛋白和组氨酸标签的编码序列。将重组质粒通过农杆菌介导的 agroinfiltration 引入到 Nicotiana benthamiana（一种野生澳大利亚烟草）的叶细胞中。通过叶提取物的荧光和 Western blot 确定，hFGF21 基因在烟草植物中正确翻译。在 agroinfection 后 7 天，重组 hFGF21 在 agroinfected 植物的叶片中积累到高达 450 μg g(-1)鲜重的水平。通过 Ni-NTA 亲和层析从植物组织中纯化重组 hFGF21，并在 3T3/L1 细胞中刺激葡萄糖摄取。这表明在 N. benthamiana 中通过 PVX 病毒载体表达的重组 hFGF21 具有生物活性。
