Animal cell nuclear protein extraction method

Abstract

The invention discloses an extraction method of nucleoprotein in animal cells. In the method, cell penetrating peptide, exogenous nucleoprotein and enhanced green fluorescent protein are utilized to jointly construct a recombinant nucleoprotein expression vector; recombinant nucleoprotein is mediated to express in nuclei; after the recombinant nucleoprotein is stably expressed in the cells, the cells are treated by using an operation solution; the cells are collected and crushed with ultrasonic wave; after the cells are centrifuged to remove precipitate, the supernatant fluid of the cells is collected, filtered and sterilized, and added into a cell culture solution; and 4 hours later, the extracted fluorescence labeled exogenous recombinant nucleoprotein enters the nuclei through cytoplasm and is positioned in the nuclei. According to the method, the extracted nucleoprotein can play functions of protein, so that morphological character of the cell changes; and results prove that the method can easily, quickly and efficiently extract the nucleoprotein with bioactivity in the cells.

Description

[technical field] [0001] The invention relates to a protein extraction method, in particular to a method for extracting an animal cell inner nucleus protein. [Background technique] [0002] The preparation of protein is a very meticulous work, involving many disciplines such as physics, chemistry and biology. There are many kinds of proteins with great differences in physical and chemical properties, so it is impossible to have a fixed procedure for the separation of various proteins. Some proteins in animal materials exist in body fluids such as blood and urine in a soluble form, and can be directly separated without extraction. However, some proteins often exist in the form of combining with other biological substances, such as nucleoproteins, which brings great difficulties to the separation work. Animal cells are composed of three parts: cell membrane, cytoplasm and nucleus. Proteins exist in large quantities in the cell membrane, cytoplasm and nucleus, and play differ...

AI Technical Summary

Problems solved by technology

However, the nuclear protein extracted by this method does not have the biological activity of nuclear protein due to protein denaturation, so it can only be used for routine analysis and detection of proteins such as Western blot, protein electrophoresis, and co-immunoprecipitation.

In addition, since the extracted protein solution contains protein denaturants, a small amount of protein denaturants in protein extracts can cause the death of cultured cells, so it cannot be added to cell culture fluids for protein functional applications

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