Multi-target-point CRISPR/Cas9 expression vector based on bacteriostasis and sterilization

Abstract

The invention discloses a multi-target-point CRISPR / Cas9 expression vector based on bacteriostasis and sterilization. The gene sequence of the multi-target-point CRISPR / Cas9 expression vector is SEQ ID NO1 and the gene sequence of a Cas9 expression vector is SEQ ID NO2. A construction method of the expression vector comprises: using a DNA segment of an oriented RNA specific recognition gene, mediating Cas9 protein to cut a DNA double strand to produce a double strand incision and break a DNA sequence, and therefore breaking the principle of DNA coding of functional protein to construct the multi-target-point CRISPR / Cas9 expression vector. Through comparison with a negative contrast, after CRISPR / Cas9 acts on DNA gyrase and dihydrofolate reductase, the survival rate of bacteria is only 10%, and the sterilization effect can reach no less than 90%.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a multi-target CRISPR / Cas9 expression vector based on antibacterial and bactericidal. Background technique [0002] Traditional antibiotics achieve the purpose of antibacterial and bactericidal by inhibiting the expression of bacterial division or metabolism-related genes. For example, penicillins and cephalosporins hinder the synthesis of bacterial cell walls; aminoglycosides and macrolides inhibit the synthesis of bacterial proteins; quinolones inhibit the replication of bacterial DNA; rifampin inhibits the synthesis of bacterial RNA; Antibiotics such as myxomycin and neomycin affect cell membrane permeability; sulfonamide antibiotics affect bacterial folic acid synthesis. Among them, bacterial DNA gyrase is the target of quinolones, and dihydrofolate reductase is the target of sulfonamide antibiotics. Under the long-term selection pressure of antibiotics, the genes e...

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Problems solved by technology

[0004] The purpose of the present invention is to provide a multi-target CRISPR / Cas9 expression vector based on antibacterial and bactericidal, aiming at solving the problem of the lack of effective theoretical guidance on the bactericidal effect of CRISPR / Cas9 on DNA gyrase and dihydrofolate, reducing the The production of medicinal properties, the problem of achieving the effect of antibacterial and bactericidal

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