Method for producing membrane protein by using stable cell strain

Abstract

The invention discloses a method for producing membrane protein by using a stable cell strain. The method mainly comprises construction of a recombinant plasmid, culture of a high expression cell strain and extraction and purification of the membrane protein. The method uses a recombinant lentiviral vector to integrate a membrane protein gene into the chromosome of the cell, so that the membrane protein gene can be continuously and stably expressed in the cell, thereby being beneficial to large-scale culture in the later stage, and solving the effect of over-expression of the membrane proteinon the production of the cell membrane protein in the prior art. At the same time, the screened high expression cell strain is cultured through a special cell culture device. The cell culture device is configured to prevent the impurities in the air from being introduced into a cell culture tank or contaminating the collected samples by arranging a multi-layered cell culture tank via sterile sampling, and is suitable for long-term and frequent sample collection and large-scale cell culture.

Description

technical field [0001] The invention relates to the field of medical molecular biology, and specifically provides a method for producing membrane protein by using a stable cell line. Background technique [0002] Membrane proteins play important roles in cell-to-cell contact, surface recognition, signal transduction, and material transport. Because of its important role, it becomes an ideal drug target. About 30% of the genes in the genome sequence encode membrane proteins, and 50% of them are the targets of currently known drugs. However, because the content of membrane protein in natural organisms is very low, and the overexpression of membrane protein is toxic to cells, and contains one or more strongly hydrophobic transmembrane regions, it is easy to form inclusion bodies during expression, which is a membrane protein The biggest bottleneck in the protein production process, so it is difficult to obtain a large number of active and soluble membrane proteins that can be ...

AI Technical Summary

Problems solved by technology

However, because the content of membrane proteins in natural organisms is very low, and the overexpression of membrane proteins is toxic to cells, and contains one or more strongly hydrophobic transmembrane regions, it is easy to form inclusion bodies during expression, which is a membrane The biggest bottleneck in the protein production process, so it is difficult to obtain a large number of active and soluble membrane proteins that can be used for research using conventional protein expression methods

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