Expression and purification system using recombinant protein as label

Abstract

The invention belongs to the technical field of biology and relates to a protein expression and purification system, in particular to an expression and purification system using a recombinant protein as a label. The protein expression and purification system is characterized in that the N end of a target protein is connected with a novel fluorescent protein sGFP (Green Fluorescent Protein) label, and the C end of the target protein is connected with histidine His peptide and is added with a polynucleotidase cutting site, thereby the protein is fused and expressed in full length; and meanwhile, a protease cutting site can be added between the target protein and the label as required. The protein expression efficiency of the system can be improved so that the yield of the target protein is improved, and the full-length protein is green in visible light so that the protein expression effect and a protein purification process can be monitored; and people can monitor the expression efficiency, the breaking degree and the purification process of the protein through naked eyes.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a protein expression and purification system, in particular to an expression and purification system with a recombinant protein as a tag. The expression and purification system can improve the yield of a target protein, monitor the protein expression effect, and perform a purification process. Background technique [0002] It is known that the production, separation and purification of proteins play an important role in life science research and production of peptide drugs. It is one of the research priorities of researchers in this field to find a good method to increase the yield of protein and make the production process and purification process visual and controllable. At present, the tags used in mature prokaryotic expression systems include maltose-binding protein tag (MBP), glutathione transferase (GST) tag, histidine His tag, etc. The folding aspect shows some role. The main ...

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Problems solved by technology

[0003] However, practice shows that the current technology still has the following problems: it is not possible to have an intuitive understanding of whether the protein is expressed, how much it is expressed, whether it has formed inclusion body precipitation, whether there is a broken expression, etc.; The direction of the protein is unknown, and the only way to judge whether there is protein flow is from the peak value; the whole process is completed in a black box, which requires a lot of financial and material resources to test, etc.

Therefore, the tags used in the protein purification system of the prior art cannot indicate the status of the target protein in expression and purification in real time, and the protein purification experiment cycle is long and the controllability is weak

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