Multiple genes deletion based on Cre-loxP marker-less gene deletion system for the strains from the genus of Pectobacterium

文献类型： 外文期刊

第一作者： Che, Shu

作者： Che, Shu;Zhuo, Yuanxu;Yang, Liping;Wang, Huan;Fan, Jiaqin;Che, Shu;Yang, Liping;Wang, Huan;Cui, Zhongli

作者机构：

关键词： Antibiotic marker; Cre-loxP multiple genes deletion; Pectobacterium

期刊名称：BIOTECHNOLOGY LETTERS （ 影响因子：2.0； 五年影响因子：2.4 ）

ISSN： 0141-5492

年卷期： 2024 年

页码：

收录情况： SCI

摘要： Objective To introduce the Cre-loxP system for constructing marker-less multiple-gene deletion mutants in Pectobacterium, overcoming limitations of antibiotic markers and enhancing the understanding of pathogenic mechanisms. Results Firstly, a plasmid named pEX18-Cre, containing a sacB sucrose suicide gene, was constructed to express Cre recombinase in Pectobacterium. Secondly, a mutant in which the loxP-Km fragment replaced the target gene was obtained through homologous recombination double-crossover with the chromosome. Finally, pEX18-Cre was introduced into the mutant to excise the DNA between the loxP sites, thereby removing the markers and achieving multiple gene deletions. By utilizing the Cre-loxP system, we successfully constructed multiple marker-less gene deletion mutants in Pectobacterium strains. Conclusions The Cre-loxP system efficiently creates marker-less multiple-gene deletion mutants, enhancing the study of Pectobacterium pathogenic mechanisms by overcoming antibiotic marker limitations.

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