Yang Jing, Qiang Weidong, Ren Suping, Yi Shanyong, Li Jian, Guan Lili, Du Linna, Guo Yongxin, Hu Huilong, Li Haiyan, Li Xiaokun
Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun 130118, China; College of Life Sciences, Jilin Agricultural University, Changchun 130118, Jilin, China.
College of Life Sciences, Jilin Agricultural University, Changchun 130118, Jilin, China.
Gene. 2018 Jan 10;639:69-76. doi: 10.1016/j.gene.2017.09.064. Epub 2017 Sep 29.
Basic fibroblast growth factor (bFGF) is a member of the fibroblast growth factors family. It is a highly specific mitogenic factor for many cell types, as though it be involved in wound repair, angiogenesis, nerve nutrition and embryonic development etc. Oil bodies have been applied for medicine, foodstuff and industry field. The heterogonous proteins expressed in oil bodies have distinct advantages, such as less purification steps and low costs. In this study, bFGF was expressed in A. thaliana seeds using oleosin fusion technology. The pOTB-bFGF vector contained an oleosin-bFGF fusion gene and a glufosinate resistance gene for selection. Transgenic A. thaliana lines were obtained by the floral dip method and protein expression was identified by SDS-PAGE and western blotting in transgenic A. thaliana lines. Moreover, MTT assays showed that the oil bodies expressed oleosin-bFGF fusion protein had a remarkable proliferation effect on NIH/3T3 cells and animal experiments showed that it could effectively decrease wound size and accelerate granulation tissue maturation. In conclusion, this may be a better method of producing oleosin-bFGF fusion protein to meet the increasing demand in its pharmacological application.
碱性成纤维细胞生长因子（bFGF）是成纤维细胞生长因子家族的一员。它是许多细胞类型的高度特异性促有丝分裂因子，似乎参与伤口修复、血管生成、神经营养和胚胎发育等过程。油体已应用于医药、食品和工业领域。在油体中表达的异源蛋白具有明显优势，如纯化步骤少、成本低。在本研究中，利用油质蛋白融合技术在拟南芥种子中表达bFGF。pOTB - bFGF载体包含一个油质蛋白 - bFGF融合基因和一个用于筛选的草铵膦抗性基因。通过花浸法获得转基因拟南芥株系，并通过SDS - PAGE和蛋白质免疫印迹法在转基因拟南芥株系中鉴定蛋白质表达。此外，MTT试验表明，油体表达的油质蛋白 - bFGF融合蛋白对NIH/3T3细胞具有显著的增殖作用，动物实验表明它能有效减小伤口大小并加速肉芽组织成熟。总之，这可能是一种更好的生产油质蛋白 - bFGF融合蛋白的方法，以满足其在药理学应用中不断增长的需求。
