Construction and analysis of pathogenicity island deletion mutants of Erwinia amylovora.
Construction and analysis of pathogenicity island deletion mutants of Erwinia amylovora.
复制标题
梨火疫病菌致病性岛缺失突变体的构建与分析
DOI：
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发表时间：
2009
期刊：
影响因子：
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通讯作者：
Dongping Wang
中科院分区：
文献类型：
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作者：
Youfu Zhao;G. Sundin;Dongping Wang
基于PCR的染色体基因一步失活技术是一种简单的基因敲除技术，广泛应用于大肠杆菌和相关肠杆菌中构建突变体。在这项研究中，我们采用这种技术来构建基因组岛和大操纵子缺失突变体的欧文氏菌amylovora，包括33.4 kb的hrp-type III分泌（T3 SS）致病岛（PAI 1）和15.8 kb的amylovoran生物合成（AMS）操纵子。两个新的T3 SS致病岛（PAI 2和PAI 3）和操纵子编码的II型分泌系统（T2 SS）的缺失表明，这些决定因素不参与植物的毒力。共接种实验表明hrp-T3 SS和AMS缺失突变体可以相互补充。这些结果进一步证实了一步失活技术可用于在大肠杆菌中产生大的缺失。食淀粉的。
An easy gene-knockout technique, PCR-based one-step inactivation of chromosomal genes, is widely used in Escherichia coli and related enterobacteria to construct mutants. In this study, we adapted this technique to construct genomic island and large operon deletion mutants of Erwinia amylovora, including the 33.4 kb hrp-type III secretion (T3SS) pathogenicity island (PAI1) and the 15.8 kb amylovoran biosynthesis (AMS) operon. Deletion of 2 novel T3SS pathogenicity islands (PAI2 and PAI3) and an operon encoding a type II secretion system (T2SS) demonstrated that these determinants are not involved in virulence in plants. Co-inoculation experiments demonstrated that the hrp-T3SS and AMS deletion mutants could complement each other. These results further confirmed that the one-step inactivation technique can be used to generate large deletions in E. amylovora.