Method for preparing soluble HIV-1 integrase recombinant protein

Abstract

The invention discloses a method for preparing a soluble HIV-1 integrase recombinant protein, and belongs to the technical field of biology. The preparation method comprises the steps of induced expression, bacteria disruption, purification and the like. The adopted expression medium contains 15 to 35 g / L of tryptone, 10 to 20 g / L of yeast extract, 2 to 3 g / L of sodium chloride, 1 g / L of glucose, 10 to 15 g / L of glycerin, 2.05 g / L of disodium hydrogen phosphate, 1.27 g / L of sodium dihydrogen phosphate, 50 mg / L of magnesium sulfate, and 60 mg / L of kanamycin; and the adopted cracking buffer solution contains 20 mM of 4-hydroxyethyl piperazine ethanesulfonic acid, 1 M of sodium chloride, 2 mM of beta-mercaptoethanol, 0.3 mg / ml of lysozyme and 5 mM of imidazole, and the pH value of the buffer solution is 9.0. The expression medium can ensure normal growth of host bacteria and efficient expression of the objective protein; and the cracking buffer solution can enhance the disruption effect of bacteria, improve the solubility of the objective protein and finally improve the yield of the objective protein.

Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of soluble HIV-1 integrase recombinant protein. Background technique

[0002] HIV-1 integrase is encoded by the HIV-1 pol gene, contains 288 amino acid residues, and has a molecular weight of about 32 kDa. Integrase catalyzes the integration of HIV-1 cDNA formed by reverse transcription into the genome of host cells, which is an essential link in the HIV-1 replication cycle. Since there are no functional analogues of integrase in human cells, inhibitors acting on integrase have little potential side effects on the human body, so integrase is considered to be an ideal target for anti-HIV-1 drug research.

[0003] In the mechanism and drug research related to HIV-1 integrase, it is usually necessary to prepare recombinant integrase protein by means of prokaryotic expression and purification. However, when using existing prokaryotic expression and puri...

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