人bFGF基因转化大豆毛状根表达体系的实验研究

发布时间：2018-07-02 20:42

本文选题：人碱性成纤维细胞生长因子(hbFGF) + 大豆　；参考：《吉林大学》2007年博士论文

【摘要】： 本研究利用基因重组技术在大豆毛状根中表达人bFGF。将人bFGF基因克隆到质粒pCAMBIA1301载体上，以大豆子叶节和胚轴为外植体，通过发根农杆菌C58C1介导转入到大豆中，，经潮霉素抗性筛选，获得潮霉素抗性的毛状根。PCR法扩增检测人bFGF基因的整合。提取毛状根中蛋白，用Western blot印迹分析人bFGF的表达。阳性毛状根中整合并表达人bFGF基因。SDS-PAGE显示样品相对分子量约为18kDa，与人bFGF标准品相对分子量一致，证明人bFGF基因在大豆的毛状根中正确表达。对转化毛状根规模化悬浮培养条件进行了优化，确定了最佳的培养条件，为利用大豆毛状根作为生物反应器工业化生产天然药物奠定实验基础。
[Abstract]:The aim of this study was to express human bFGF in soybean hairy roots by gene recombination technique. The human bFGF gene was cloned into plasmid pCAMBIA1301 and transformed into soybean by Agrobacterium tumefaciens C58C1, and was screened by hygromycin resistance. Hygromycin resistant hairy roots were amplified by PCR to detect the integration of human bFGF gene. The protein was extracted from hairy root and the expression of human bFGF was analyzed by Western blot. The expression of human bFGF gene. SDS-PAGE in the positive hairy root showed that the relative molecular weight of the sample was about 18kDa. it was proved that the human bFGF gene was correctly expressed in the hairy root of soybean. The optimal culture conditions of the transformed hairy roots were optimized and the optimum culture conditions were determined, which laid the experimental foundation for the industrialized production of natural drugs by using soybean hairy roots as bioreactor.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2007
【分类号】：R346

【引证文献】