Preparation and renaturation method of chicken interferon gamma

Abstract

Provided is a method for preparing and renaturing chicken interferon ?, comprising the following steps: (1) performing PCR amplification on the genes of chicken interferon ?; (2) subcloning the product of step (1) to a pET32a expression vector having a 6-His tag; (3) transforming the product of step (2) into cells of Escherichia coli M15; (4) performing induction and expression on the product of step (3), then cracking to acquire a protein expression product; (5) purifying the expression product of step (4); and (6) renaturing recombinant protein purified to acquire a product. Employment of the pET32a and of Escherichia coli M15 allows for the expression product of chicken interferon ? gene to reach after purification a purity of 95%, which is greater than the 85% purity of a conventional method, and for the renaturation to reach a success rate between 38% and 39%, which is greater than the 20% success rate of the conventional method. Increase of active ingredient in total protein prepared improves treatment effects and reduces costs.

Description

technical field [0001] The invention belongs to the technical field of livestock and poultry medicines, in particular to a preparation and refolding method of chicken interferon gamma. Background technique [0002] In the poultry farming industry, infectious diseases caused by viruses account for about 70% of the total number of diseases. Vaccines and antibiotics are often used to prevent and control these diseases, but vaccines cannot resist the mutation of pathogens and the emergence of new diseases. Lower downside. Through research, people have developed interferon that can interfere with the replication of viruses in infected and non-infected tissues. Among them, interferon gamma (type II interferon) has a strong immune regulation ability and is the most important immune regulation factor in the body. It is manifested in the effect on the activity of host immune cells, which can increase the expression of MHC class II molecules on the surface of macrophages and enhance ...

AI Technical Summary

Problems solved by technology

[0008] Interferon is a small molecular protein, which has disadvantages such as unstable conformation and easy overexpression and aggregation into inclusion bodies during the expression process. When the above method is used, the expression product is likely to appear in the form of tight inclusion bodies, which is not conducive to purification and complexation. Sex, unfavorable for the spatial conformation of interferon

[0009] High concentrations of denaturants need to be introduced in the purification process of expression products, the purity of purified products is not high (about 85%), the renaturation process is cumbersome, the success rate of renaturation between batches is low, and the biological activity of renaturation products is low, which leads to the final medicine The cost is extremely high

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