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通过构建重组表达质粒载体p139035S-KGF1和根癌农杆菌介导在红花(Carthamus tinctorius)中表达角质细胞生长因子(KGF-1)。从侵染到诱导生根共需要14周,转化率达0.1%。红花子叶在潮霉素筛选培养基上培养4-5周后便可获得丛生芽,再生芽移入含潮霉素的伸长生根培养基,培养4-8周可诱导生根。通过PCR、Southern blot、RT-PCR及Western blot检测证明目的基因KGF-1已经整合到红花细胞的染色体中,实现了KGF-1外源蛋白在红花中的成功表达,为开发KGF-1蛋白新的生产途径奠定了基础。
Expression of keratinocyte growth factor (KGF-1) in carthamus tinctorius was mediated by the construction of the recombinant expression plasmid vector p139035S-KGF1 and Agrobacterium tumefaciens. It takes 14 weeks from infection to induction of rooting, with a conversion rate of 0.1%. Crocus buds can be obtained after cultivating safflower cotyledons on hygromycin selection medium for 4-5 weeks, and regeneration buds are transferred to elongation rooting medium containing hygromycin, and rooting can be induced after 4-8 weeks of culture. The results of PCR, Southern blot, RT-PCR and Western blot showed that the target gene KGF-1 has been integrated into the chromosome of safflower cells and successfully expressed the KGF-1 foreign protein in safflower. In order to develop KGF-1 The new ways of production of protein laid the foundation.