Method for highly efficiently deleting large fragments of genome based on endogenous CRISPR-Cas system of zymomonas mobilis and application thereof

A technology of Zymomonas and deletion method, applied in the field of efficient deletion of large genome fragments, can solve the problems of host cell toxicity and difficult application, and achieve the effect of avoiding cytotoxicity and efficient deletion of large genome fragments
CN110408642AActive Publication Date: 2019-11-05武汉睿嘉康生物科技有限公司
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
武汉睿嘉康生物科技有限公司
Publication Date
2019-11-05

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Abstract

The invention belongs to the technical field of genetic engineering, and in particular, relates to a method for highly efficiently deleting large fragments of a genome based on an endogenous CRISPR-Cas system of zymomonas mobilis and an application thereof. The method includes the steps: constructing a plasmid containing an artificial CRISPR expression unit; determining a large-fragment editing target site, selecting a guideRNA sequence for the editing target site and designing a primer; constructing the guideRNA primer sequence to the plasmid containing the artificial CRISPR expression unit,and constructing a vector; constructing a donor DNA sequence to the vector, to obtain an editing plasmid; and transforming the editing plasmid into a receptive cell, and editing. With a high-throughput gene editing platform based on the endogenous CRISPR-Cas system of zymomonas mobilis, the editing purpose of deleting the large fragments of the genome of the strain is achieved, and the developmentof metabolic engineering, system biology and synthetic biology is promoted.
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Description

technical field

[0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a method for efficiently deleting large genome fragments based on the endogenous CRISPR-Cas system of Zymomonas mobilis and its application. Background technique

[0002] In recent years, the use of microorganisms for metabolic engineering, systems biology, and synthetic biology has made good progress. For the rational design and construction of microbial cell factories, living cells or enzymes are used to treat renewable biomass, such as It provides an important theoretical basis for the transformation of cellulose and other substances, the production of bioenergy, and the industrialization of biosmelting. Bioenergy regeneration is one of the effective means to solve the problems of resource and energy shortage and serious environmental pollution that human beings are currently facing.

[0003] Because Zymomonas mobilis can produce alcohol naturally, it ...

Claims

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