Linearized DNA vector pHB-1 plasmid and test kit prepared from same and used for editing bacterial genomes

Abstract

The invention discloses a linearized DNA vector pHB-1 plasmid and a test kit prepared from the same. The genetic sequence of the linearized DNA vector pHB-1 plasmid is shown as SEQ ID NO: 1; the testkit comprises a linearized DNA vector pHB-1 plasmid, an efficient T4DNA ligase, a PCR sequencing primer tL3-seq and an experimental control substance. Wherein the experimental control comprises a specific gRNA sequence primer for editing an escherichia coli MlaC gene, a DNA repair template for introducing MlaC L11R point mutation and a PCR sequencing primer for confirming the MlaC L11R point mutation; the test kit disclosed by the invention can be used for carrying out gene editing experiments on various bacteria, has repeatability and stability, and can be used for editing one or more targetgenes, and the editing efficiency can reach 90-100%.

Description

technical field

[0001] The invention relates to the technical field of genetic engineering, and belongs to the field of high-end biotechnology, in particular to a linearized DNA carrier pHB-1 plasmid and a kit for editing bacterial genomes prepared therewith. Background technique

[0002] Bacterial genome editing is the purpose of studying its functions and traits by interrupting, deleting or replacing target genes on the genome. It plays an important role in gene editing and its application can be extended to engineering bacteria transformation, antibiotic research and development, and resistance Drug mechanism research and many other aspects. Although traditional plasmid transformation methods can be used to achieve transformation in a variety of bacteria, it is time-consuming and labor-intensive, and cannot precisely control the genome transformation position and transformation sequence, and large-scale sequencing and repeated induction are required for a transformed micr...

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