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[目的]构建银杏GGPPS转运肽与GFP融合基因表达载体。[方法]以银杏为材料,采用DNA重组技术克隆GGPPS基因质体转运肽(TP)序列,并将其与高效植物表达载体p1304+连接形成融合表达载体(p1304+-TP);冻融法转化根瘤农杆菌EHA105,构建工程菌(EHA105-p1304+-TP)。[结果]成功构建了银杏GGPPS转运肽与GFP融合基因表达载体及农杆菌工程菌。[结论]为进一步研究TP转运肽的亚细胞定位奠定基础,有助于阐明银杏内酯前体生物合成关键步骤的分子机理,同时为银杏内酯的代谢工程研究提供重要依据。
[Objective] To construct Ginkgo GGPPS transit peptide and GFP fusion gene expression vector. [Method] With Ginkgo biloba as material, the GGPPS gene transfer peptide (TP) was cloned by DNA recombination and ligated with p1304 + to construct the fusion expression vector (p1304 + -TP) Bacillus EHA105, construction of engineered bacteria (EHA105-p1304 + -TP). [Result] The GGPPS transit peptide and GFP fusion gene expression vector and Agrobacterium engineering bacteria were successfully constructed. [Conclusion] This study may lay the foundation for the further study on the subcellular localization of TP transit peptide and help elucidate the molecular mechanism of the key steps of ginkgolide precursor biosynthesis, and provide an important basis for the metabolic engineering research of ginkgolide.