Preparation method of human papillomavirus and heat shock protein recombinant protein

Abstract

The present invention provides a preparation method of a human papillomavirus and heat shock protein recombinant protein. The preparation method comprises the following steps: A, construction of engineering bacteria expressing human papillomavirus and heat shock protein recombinant protein is conducted; and B, inducing expression and target protein purification are conducted; wherein the engineered bacteria are obtained by codon-optimizing a coding gene of the recombinant protein, then constructing into pET28a plasmids and transferring into escherichia coli. The provided optimized gene of thehuman papillomavirus and heat shock protein recombinant protein can be highly expressed, and an expression level of the target protein accounts for 20% of whole bacterial proteins. In the process of protein purification, inclusion body is dissolved by sodium lauryl sarcosinate and subjected to renaturation on a combination column, and the method effectively avoids complicated processes, reduces uses of a denaturing agent urea or guanidine hydrochloride, reduces waste liquid discharge, has many advantages of low cost, high renaturation rate, convenient operation, etc., and provides a new idea for preparation of the human papillomavirus and heat shock protein recombinant protein.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a preparation method of human papilloma virus and heat shock protein recombinant protein. Background technique [0002] Among gynecological tumors, the incidence of cervical cancer is second only to breast cancer, ranking second. About 200,000 women worldwide die from this disease every year. Statistics in recent years show that there are about 131,500 new cervical cancer patients in my country every year, accounting for 28% of the total number of new cervical cancer cases in the world. About 290,000 women die of cervical cancer every year in the world, of which about 50,000 are in my country. , The rising trend of young cervical cancer patients is obvious. According to reports, the occurrence of cervical cancer is closely related to human papillomavirus (HPV) infection, and some high-risk HPV virus infections are likely to eventually lead to cervical cancer. ...

AI Technical Summary

Problems solved by technology

[0005] At present, it is very difficult to express and purify HPV E6 from Escherichia coli (E.coli) (Two-stepchromatographic purification of glutathione S-transferase-tagged humanpapillomavirus type 16E6protein and its application for serology, ProteinExpression and Purification Volume 132, April 2017, Pages 19-26), because of its strong hydrophobicity, it is easy to produce protein aggregates, which affects subsequent purification

Therefore, when purifying HPV E6-related recombinant proteins, the research data reported at home and abroad are to design purification tags (such as GST, MBP, HIS, etc.) on related recombinant proteins, and use affinity chromatography to purify to obtain high-purity target proteins. For example, the E. coli expression system expresses human papillomavirus (HPV) type 16 E6E7HSP65 fusion protein using HIS tag purification, and the redundant tag (redundant amino acid) of the final target protein is not removed, although the redundant amino acid cannot be seen in the short term Side effects, but the safety of long-term use cannot be guaranteed, and it will affect the approval of new drugs

Images