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表皮生长因子(EGF)具有促进多种细胞增殖的作用, 尤其在维持消化道黏膜完整和促进消化道溃疡愈合方面作用巨大。以前的研究多集中在细菌和酵母中表达, 本研究试图以番茄作为生物反应器生产重组人 EGF, 使之成本降低, 应用方便。依据人 EGF 的基因序列, 设计合成了番茄密码子偏爱的人 EGF 基因, 并将其构建到植物表达载体 pCAMBIA2300 中, 通过农杆菌介导得到了含有人 EGF 基因的转基因番茄。放射免疫法检测到每克鲜重果实中的表达量达 3.48 ± 1.01 ng。将果汁灌喂小白鼠 15 天(相当于每鼠每天喂服 24 ng rhEGF)能显著抵抗酒精引起的胃溃疡形成, 溃疡指数由 42.20 ± 18.13 下降为 16.25 ± 9.57。
Epidermal growth factor (EGF) has the effect of promoting a variety of cell proliferation, especially in the maintenance of gastrointestinal mucosal integrity and promote the healing of peptic ulcer disease. Previous studies focused on bacterial and yeast expression. In this study, tomato was used as a bioreactor to produce recombinant human EGF, which reduced the cost and facilitated the application. Based on the human EGF gene sequence, we designed and synthesized a human EGF gene, which is the preferred codon for tomato, and constructed it into the plant expression vector pCAMBIA2300. The transgenic tomato containing EGF gene was obtained by Agrobacterium tumefaciens. Radioimmunoassay showed that the expression level of fresh fruit per gram was 3.48 ± 1.01 ng. The mice fed the juice for 15 days (equivalent to 24 ng rhEGF per day) were significantly more resistant to alcohol-induced gastric ulcer formation, with the ulcer index decreasing from 42.20 ± 18.13 to 16.25 ± 9.57.