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
作者： Che, Shu 1 ; Zhuo, Yuanxu 1 ; Yang, Liping 1 ; Wang, Huan 1 ; Cui, Zhongli 5 ; Fan, Jiaqin 1 ;
作者机构： 1.Nanjing Agr Univ, Dept Plant Pathol, Lab Bacteriol, Nanjing 210095, Peoples R China
2.Jiangxi Acad Agr Sci, Plant Protect Inst, Nanchang 330200, Peoples R China
3.Yunnan Acad Agr Sci, Inst Grain Prod, Kunming 650205, Peoples R China
4.Inst Agr Sci Suzhou, Suzhou 215155, Peoples R China
5.Nanjing Agr Univ, Key Lab Agr Environm Microbiol, Minist Agr & Rural Affairs, Coll Life Sci, Nanjing 210095, Peoples R China
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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