A kind of preparation refolding method of chicken interferon gamma

Abstract

The invention relates to a preparation and renaturation method of chicken interferon gamma, which comprises the following steps: (1) performing polymerase chain reaction (PCR) amplification on chicken interferon gamma; (2) subcloning the product obtained by the step (1) onto a pE32a expression vector having a 6His mark; (3) transferring the product obtained by the step (2) into a cell of Escherichia coli M15; (4) inducing and expressing the product obtained by the step (3), and splitting to obtain the product of expression; (5) purifying the product of expression, which is obtained by the step (4); and (6) renaturing the purified recombinant protein to obtain the finished product. In the invention, by using the Pet32a and the Escherichia coli M15, the purity of the purified product of thehigh-efficiency expression of a chicken interferon gamma gene reaches 95 percent, which is higher than the 85 percent of the conventional method, the renatured protein yield reaches 95mu g / ml, which is higher than the 30 mu g / ml of the conventional method, and the renaturation success rate is up to 38 to 39 percent, which is higher than the 20 percent of the conventional method; and the content of the active ingredients in the prepared total proteins is increased, the treatment effect is enhanced, and the cost is lowered.

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

[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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