Transformation Of Large DNA Fragment Of Panax Ginseng(100Kb)into Ganoderma Lucidum

The Panax ginseng C.A.Meyer belongs to Araliaceaae Panax plant. The dired roots and rhizome of ginseng is a treasure medicine in China, which is called the King of Herb. It acts as an excellent nourishing five-orgen, calming nerves, stopping palpitates with fear, improving eyesight. complex chemical constituents of ginseng, ginsenoside is considered to be the main active ingredient of ginseng.Ganoderma lucidum (Curtis.) P. Karst. belongs to Large filamentous fungi of the Ganoderma lucidum genus, which is treasures medicinal fungi food and it is well known as grass jelly in China. Ganoderma lucidum acid is the main chemical constituents of Ganoderma lucidum, it can protect the liver, anti-inflammatory and other functions.Ganoderma lucidum acid synthesis pathway have similar biosynthetic pathway with the ginsenosides, it is MVA biosynthetic pathway. so select Ganoderma lucidum as a receptor, transformed a100kb DN A fragment of Panax ginseng into Ganoderma lucidum protoplasts by electroporation, to disscus the feasibility of transforming large DNA fragments into Ganoderma lucidum, providing theoretical and technical for Ganoderma lucidum resource, and lay the foundation for the heterologous production of ginsenoside, provide a new technology system for analysising the function of ginseng DNA fragment.The following results were obtained:1The optimum conditions for the protoplast regeneration are:strain age is5d, the enzymatic hydrolysis time is2.5h, the temperature is31℃, pH5.5, lywallzyme concentration is2.0%, osmotic stabilizer is0.6mol/mL mannitol the solid MYG is used as regeneration medium,2The optimum conditions for the transformation are:a number of regenerative clones were obtained on the selective medium when using4mm spaced electrodes at250volts. The PCR results indicated that the ginseng large DNA fragment had been transformed into the Ganoderma lucidum.3Through the determination of growth state and curve, showed that the transformant growed slowly.4Ferment Transformants strains and non-transformed strains, the content of Ganoderma lucidum acid showed that,:the resistant transformants than non-transformed strains decreased.