| Fungi polysaccharides as widespread biological macromolecular material nature,have efficient immune enhancement effect,is widely applied in regulating immunity,anti-tumor,antioxidant and so on health care.Fungi,however,is widespread in all kinds of fungi polysaccharide content is very low,the industrial production of low cost and high yield,it is hard to meet the demand of the market.Using genetic breeding means breeding mushroom polysaccharides high yield strains can provide ideas for all kinds of fungi polysaccharide yield increase.This article build mushroom polysaccharide yield promoter in alternative technology engineering bacteria,application structure nest type of aspergillus efficient glyceraldehyde 3-phosphoric acid dehydrogenase gene promoter(GDP)of promoter alternative Pleurotus ostreatus TD 300 glucan synthesis of key enzymes of glucan synthase gene promoter,enhance glucan synthase gene expression,yield of glucan.In transformation of homologous recombination fragments using liposome mediated transformation,improving the efficiency of conversion from the simplified conversion step.This method is compared with the traditional protoplast transformation,short cycle,high efficiency.This article build Pleurotus ostreatus TD 300 high-yield strain experiment achieved the following progress:(1)In terms of building segments of homologous recombination,land use-overlap PCR method,express to fusion of four segments of stay connected,save steps,experiment materials.(2)By liposome mediated method into Pleurotus ostreatus TD 300 grinding hyphae,conversion steps is convenient,the operation is simple.Compared with the protoplast transformation method to save a lot of time.(3)Glucan synthase gene promoter is replaced by the method of homologous recombination,enhancing the gene expression quantity of polysaccharide yield increase.(4)Achieved high yield strains of Pleurotus ostreatus TD 300,polysaccharide yield can be increased by 2.3 times. |
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