What is the purification method of protein purification system

Protein Purification System what is the purification method, you need to know the DNA sequence encoding the purified protein, see which cells or tissues overexpress the target gene, and design primers for the gene to amplify the arf of the target DNA fragment. This is the so-called acquisition of target gene fragments.

Constructing expression vectors: In the prokaryotic or eukaryotic expression vectors that obtain the expression characteristics of the genes themselves, the main problem in this step is to construct plasmids and expression systems for the genes of interest. Prokaryotic expression time is short, low cost, and large-scale expression are a priority; the gene is not expressed in E. coli, the problem arises of codon optimization, and in consideration of better activity and purified protein, researchers choose to express in Pichia yeast. Codon optimization is important for successful expression.

Protein Purification System what is the purification method:

1. Precipitation.

2. Electrophoresis: The charged protein is above or below its isoelectric point and can be moved to the negative electrode or the positive electrode of the electric field in the electric field. Supports include thin film electrophoresis, electrophoresis, etc.

3. Dialysis: using two dialysis bags to separate macromolecules from proteins and small molecular organic compounds.

4. Chromatography: Ion-exchange chromatography uses the free nature of proteins. Under a specific pH, proteins have different charges and properties, and can be separated by ion-exchange chromatography. In anion exchange chromatography, proteins with less negative power are eluted first. Molecular sieves, also known as gel filtration. The tiny proteins get into the pores and stay there for a long time. Larger proteins cannot enter the pores and flow directly out.

5. Protein Purification System what is the method of purification, ultracentrifugation: protein purification can be used to determine the molecular weight can be used as the protein. Proteins that form different densities are separated.