Wang Meizhen, Deng Yuxing, Shao Fenjuan, Liu Miaomiao, Pang Yongqi, Li Caili, Lu Shanfa
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Peking Union Medical College, No.151, Malianwa North Road, Haidian District, Beijing, 100193, China.
State Key Laboratory of Tree Genetics and Breeding, Research Institute of Forestry, Chinese Academy of Forestry, Beijing, China.
Methods Mol Biol. 2017;1640:173-189. doi: 10.1007/978-1-4939-7165-7_12.
Small RNA-mediated gene silencing is a vital regulatory mechanism in eukaryotes that requires ARGONAUTE (AGO) proteins. Salvia miltiorrhiza is a well-known traditional Chinese medicinal plant. Therefore, it is important to characterize S. miltiorrhiza AGO family genes as they may be involved in multiple metabolic pathways. This chapter introduces the detailed protocol for SmAGO gene prediction and molecular cloning. In addition, an Agrobacterium-mediated genetic transformation method for S. miltiorrhiza is presented. These methodologies can be used to functionally study SmAGO genes as well as other genes of interest in S. miltiorrhiza.
小RNA介导的基因沉默是真核生物中一种重要的调控机制，该过程需要AGO（精氨酸）蛋白的参与。丹参是一种著名的传统中药材。因此，对丹参AGO家族基因进行表征具有重要意义，因为它们可能参与多种代谢途径。本章介绍了丹参SmAGO基因预测和分子克隆的详细方案。此外，还介绍了一种农杆菌介导的丹参遗传转化方法。这些方法可用于对丹参SmAGO基因以及其他感兴趣的基因进行功能研究。
