| Salvia miltiorrhuza Bunge is a well-known Chinese herbal plant, and its roots,known as Danshen in Chinese, are widely used in the clinical treatment of menstrual disorders and blood circulation diseases etc. Salvia miltiorrhiza includes Liposoluble diterpenoids and water-soluble compound. Diterpenoids especially Tanshinoneâ…¡A is a kind of important medicine compound which is used to treat cardiovascular and cerebrovascular diseases.Application gateway cloning technology to build Danshen CPS gene over-expression vector and by Agrobacterium rhizogenes Danshen be hairy roots and by Agrobacterium tumefaciens Danshen and tobacco transgenic lines were obtained. To further lay the foundation for screening of high-yielding strains,for studying further function of CPS gene in the Metabolic pathway.According to the laboratory acquired CPS gene cDNA and Gateway cloning technology requirements, Two pair of primers containing attB adapter were designed separately by Gateway cloning technology.Will be to build a CPS gene vector dual pH7WG2DThrough electroporation to construct a good vector pH7WG2D-CPS into Agrobacterium rhizogenes in ACCC10060 and EHA105.By Agrobacterium tumefaciens was Danshen hairy roots and Salvia transgenic strains and tobacco transgenic strains.Through PCR, enzyme digestion and sequencing to identify vector pH7WG2D-CPS to build successful. CPS Gene was transformated into salvia miltiorrhiza via Agrobacterium-mediated Method to obtain the CPS overexpression masculine hairy root and the masculine adult plant and the tobacco masculine adult plant.Construction of vector application gateway technology is simple, rapid and high conversion rate, Agrobacterium-mediated transformation method is simple and obtained transgenic hairy roots Danshen positive and Higher conversion rate over 70%. Transgenic plants obtained in the positive rate of less than 10%. Materials obtained through positive danshen for further study functional genomics and lay the foundation for high-yielding strains. |
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