A method for the production of polyhedrons that both reduce internal virions and entrap foreign proteins

Abstract

The invention relates to the field of biotechnology, and in particular relates to a production method of a polyhedrin capable of reducing internal virus particles and embedding heterologous protein. The production method comprises the following steps: (1) constructing a pFast-PH plasmid containing a polyhedrin gene; (2) constructing a recombinant pFast-PH-pPH-ac12-egfp plasmid containing egfp andac12 genes; (3) constructing ac12 gene over-expression bacmid-bAcac12OE; (4) implementing cell transfection, so that a recombinant virus is obtained; and (5) infecting the recombinant virus with cells, and purifying the polyhedrin. According to the production method provided by the invention that a recombinant exogenous gene fragment is interpolated at the back of the polyhedrin gene, influence tothe formation of the polyhedrin is avoided, and moreover, the heterologous protein is embedded into the polyhedrin; via a method of conducting fusion expression on a baculovirus efficient and strongpromoter, namely polyhedrin promoter, and AC12 protein, high-level expression of recombinant protein is achieved and the level of the inclusion virus particles in the polyhedrin is reduced; therefore,conditions are created for further development and utilization of a baculovirus-insect expression system and for developing the polyhedrin into a novel medical carrier and a nano-material.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a production method of a polyhedron capable of reducing internal virus particles and embedding foreign proteins. Background technique [0002] Baculovirus polyhedron gene is a high-level expression protein produced by baculovirus in the late stage after infection of the host. The polyhedron promoter is a very late super-expression promoter of baculovirus, which has been successfully applied to the baculovirus expression system for high-level expression of foreign genes. Polyhedrin proteins can form ultra-macromolecular protein crystal polyhedrons after polymerization. Current research shows that polyhedrons can protect virus particles and proteins embedded in polyhedrons, allowing them to preserve their activity for a long time. However, the existence of virus particles inside polyhedrons affects the safety of polyhedrons as medical nanomaterials. It is the long-cherished wish of m...

AI Technical Summary

Problems solved by technology

However, the existence of virus particles inside polyhedrons affects the safety of polyhedrons as medical nanomaterials. It is the long-cherished wish of medical and chemical workers to reduce the virus particles contained in polyhedrons.

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