Transformation Of150kb Large DNA Fragment Of Panax Ginseng Into Ganoderma Lucidum

The Panax ginseng belongs to Araliaceaae Panax plant,one of the most important plants used in traditional medicine. Ginsenoside is one of main effective components. The scientists only have certain individual genes to carry out a more traditional molecular biology. There are few studies of ginseng genome, has yet to see the complete gene, This condition limits depth-study in breeding, important agronomic traits, and quality.Ganoderma lucidum as one kind of mushrooms, belongs to Basidiomycotav,has anticancer and other kinds of activity. Ganoderma acid is one of the main effective components. Ganoderma lucidum acid metabolic pathways of metabolic pathways has similar with the ginsenoside MVA biosynthetic pathway, and it has small genomic,easily to be translated,short growth cycle and some other advantages.So the experiment used the BIBAC of The Panax ginseng which had been sed up,choosed Ganoderma lucidum as transforming receptor then transformed the large DNA fragment(150kb) into the Ganoderma lucidum protoplast by electroporation. Try to evidence the function of genes which contained in the large segment of ginseng DNA by Ganoderma Acid fermentation, lay theorical and technological foundation for germplasm creation of Lucid Ganoderma, breeding and industrialized production of active ingredient.Get the results as followed:1. The optimal conditions of Lucid Ganoderma protoplast isolation and regeneration: spawn age is5days, the enzymatic hydrolysis time is2.5hours,the temperature is31℃,the hydrolysis pH value is5.5, the osmotic pressure stabilizer selects mannitol. Mycelium grows fastest when the regenerative form is double layer culture method,The optimal medium for protoplast regeneration is MYG solid medium.2. The optimal conditions of transformation of electricity:electrode gap is4mm, voltage strength is200V,quantity addition of protoplast is60μL, quantity addition of large DNA fragment is5μL3.Analyses of the transgenic with PCR test,showed that large DNA fragment had been transformated into Ganoderma lucidum.4. Through the determination of growth curve,showed that the transformant growed slowly.5. Through the determination of the content of the Ganoderma acid,showed that the content of the transformant is a little lower.