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Method for gene editing in amycolatopsis orientalis

The invention discloses a method for performing gene editing in amycolatopsis orientalis. The method comprises the following steps: (1) constructing upstream and downstream homologous arm fragments nrps8-arm-A and nrps8-arm-Z of a target gene nrps8 sequence for DNA repair after knockout; (2) constructing a DNA (Deoxyribose Nucleic Acid) recombinant fragment gDNA-nrps8 capable of coding the sgRNA; (3) constructing a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas9 editing plasmid pLYCS01; and (4) electrically transforming the pLYCS01 and the gDNA (deoxyribonucleic acid)-nrps8 into the A.keratiniphila cell, so as to obtain the A.keratiniphila cell. The invention also discloses an application of the nrps8 gene of the A. keratiniphila as a gene knockout target of CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / CRISPR associated protein 9) in the preparation of vancomycin.
Owner:SHANGHAI JIAOTONG UNIV