Method for efficiently expressing active Dog Growth Hormone (GH) growth hormone

Abstract

The invention discloses a method for high-efficiency expression of active Dog Growth Hormone (GH) growth hormone, and relates to the technical field of gene recombination, and the method comprises the following steps: step 1, synthesis of a target gene, step 2, sequencing of the target gene, step 3, connection of a Growth Hormone (GH) gene and an expression vector, step 4, transformation of a recombinant plasmid PTZY1-GH, step 5, enzyme digestion identification of the recombinant plasmid, and step 6, purification of the recombinant plasmid. The method comprises the following steps of: 1, extracting an endotoxin-free plasmid, 7, transfecting CHOS (hamster ovary) cells, 8, purifying target protein, and 9, verifying whether the expressed target protein has disulfide bonds or not and verifying the proliferation activity of the cells. The expressed protein is stable in quality and high in yield which can reach 1-2 g / L, the product is uniform, a domestic culture medium is adopted for cell culture, compared with an imported culture medium, the raw material cost can be greatly saved, large-scale production is easy, and meanwhile, the recombinant Dog Gh recombinant protein has Nb2-11 cell proliferation activity and can be used for follow-up development of dog growth hormone drugs.

Description

technical field [0001] The invention relates to the technical field of gene recombination, in particular to a method for efficiently expressing active Dog Growth Hormone (GH) growth hormone. Background technique [0002] Recombinant protein technology uses the method of genetic engineering to connect the target protein DNA obtained in vitro to an expression vector, and the constructed expression vector is transfected into the target cell, and the target protein is obtained by in vitro culture. Production is mainly divided into two categories: eukaryotic expression system and prokaryotic expression system: the prokaryotic expression system mainly uses Escherichia coli E Coli as an expression strain, which has the advantages of low cost, high efficiency, and low environmental requirements. The disadvantage is that the expression Proteins often exist in the form of inclusion bodies, which cannot fold normally and present a correct three-dimensional structure, resulting in the i...

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Problems solved by technology

[0004] The invention provides a method for efficiently expressing active Dog Growth Hormone (GH) growth hormone, which solves the problem of slow growth of dogs suffering from dwarfism through injection of growth hormone, but currently the growth hormone injected into dogs is bovine The source of growth hormone has poor effect, and the Escherichia coli expression system has low yield and poor activity, and the growth hormone used in dogs has poor growth stability, low safety and high cost.

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