23 Jul Taxodium Zhongshanha 406 mesophyll protoplast separation, purification and high-efficiency conversion method
Abstract
The invention discloses a Taxodium Zhongshanha 406 mesophyll protoplast separation, purification and high-efficiency conversion method. The method comprises the following steps: 1, carrying out dark pretreatment on a material tender leaves of 30 days-old good-growing tissue culture seedlings of Taxodium Zhongshanha; 2, separating and purifying mesophyll protoplasts; 3, extracting 50-80 [mu]g of a plasmid, which is a vector p2FGW7.0, from 15-20 mL of an overnight cultured bacterium liquid; and 4, converting the protoplasts through PEG mediation. The Taxodium Zhongshanha 406 mesophyll protoplast separation, purification and high-efficiency conversion method has the advantages of simplicity and easiness in operation, obtaining of massive, complete and pure Taxodium Zhongshanha mesophyll protoplasts, and high plasmid conversion rate reaching 80%. The method solves a problem that the gene functions of the Taxodium Zhongshanha cannot be deeply researched, provides a certain reference for the establishment of other coniferous species protoplast instantaneous expression systems, and has great application values in the field of forest tree gene engineering.
Year: 2017
Country: CN
Doc No: 107267549