Method for tissue culturing ice edge plant in high mountains and for production of excreting protein-from cold induction

Abstract

A method for the tissue culture of alpine perglacial plant and preparing the cold-induced secretoprotein includes sampling, determining the candidate standard, screening the plant, tissue culture of the leaf coming from the apline periglacial plant, pretreating by hormone GA3 for de-differentiating, enlargig reproduction, differentiating again, evaluating its variation, cold inducing by shake culture of suspended tissue, testing the secretoprotein, and separating the protein.

Description

technical field [0001] The invention belongs to protein engineering technology, in particular to a method for alpine periglacial plant tissue culture and cold-induced secretion protein production. Background technique [0002] Plant secreted protein refers to the protein secreted by plant cells into the apoplast, which can be secreted into the culture medium during liquid shaking culture. Common plant secreted proteins include signal polypeptides or antifreeze proteins. For example, calmodulin is a signal polypeptide. It has been confirmed by more than 10 years of research at home and abroad that calmodulin not only exists outside the plant cells, but also has the ability to promote the proliferation of suspension culture cells and their protoplasts and promote the growth of secondary cells in the extracellular site. Phosphoribulose carboxylase / oxygenase small subunit gene light-independent expression and other important biological functions; the research on antifreeze prot...

AI Technical Summary

Problems solved by technology

Due to the non-universal distribution of antifreeze protein, it is difficult to find it from several specific plants, and the traditional apoplast extraction technology is difficult to master, which greatly limits the isolation of antifreeze protein secreted by cold induction

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