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为探索大豆油体蛋白与人bFGF(碱性成纤维细胞生长因子)蛋白在植物油体中融合表达的可行性,以大豆总DNA为模板,通过PCR扩增得到大豆油体蛋白基因(Ddoil)及启动子(Ddprm),构建Ddprm启动的Ddoil基因与植物偏好密码子改造的bFGF基因融合表达载体p1390Ddprm-Ddoil-bFGF,花侵染法转化拟南芥,对T2拟南芥抗性植株基因组进行PCR检测,对种子总蛋白进行SDS-PAGE分析、Western blot检测、ELISA检测及细胞活性测定,为bFGF在其他油料作物的油体系统的表达提供参考。
In order to explore the feasibility of fusion expression of soybean oil body protein and human bFGF (bFGF) protein in plant oil, total soybean DNA was used as template to amplify soybean oil body protein gene (Ddoil) and (Ddprm) was used to construct Ddprm-initiated Ddoil gene and plant-preferred codon modified bFGF gene fusion expression vector p1390Ddprm-Ddoil-bFGF. The Arabidopsis thaliana was transformed by flow cytometry. The genome of T2-resistant Arabidopsis thaliana was PCR- SDS-PAGE, Western blot, ELISA and cell viability assay were performed on the total protein of seed to provide a reference for the expression of bFGF in the oil system of other oilseed crops.