An improved, versatile and efficient modular plasmid assembly system for expression analyses of genes in Xanthomonas oryzae.
An improved, versatile and efficient modular plasmid assembly system for expression analyses of genes in Xanthomonas oryzae.
复制标题
一种改进的、多功能的、高效的模块化质粒组装系统，用于米黄单胞菌基因的表达分析
DOI：
10.1111/mpp.13033
复制
发表时间：
2021-04
影响因子：
4.9
通讯作者：
Chen G
中科院分区：
文献类型：
--
作者：
Zou L;Zhang C;Li Y;Yang X;Wang Y;Yan Y;Yang R;Huang M;Haq F;Yang CH;Chen G
水稻黄单胞菌致病变种稻生黄单胞菌（Xoo）和水稻条斑病菌（Xoc）分别侵染水稻，引起水稻白叶枯病和水稻细菌性条斑病，这是稻田中两种具有重要经济影响的细菌性病害。Xoo和Xoc与水稻的相互作用可作为研究细菌致病性和宿主组织特异性基本方面的模型。然而，由于一些能在水稻黄单胞菌致病变种中稳定复制的广宿主范围载体是低拷贝数质粒，所以Xoo和Xoc需要一种改进的用于基因表达分析的载体系统。为克服这一局限，我们开发了一种模块化质粒组装系统，以便将功能性DNA模块从入门载体以高拷贝数的方式转移到源自pHM1的骨架载体中。我们证明了我们的载体系统在实验室条件下以及与宿主水稻和非宿主烟草细胞相关联时用于蛋白质检测以及毒力基因表达定量的可行性。该系统还允许进行等同于单拷贝染色体整合系统的突变体互补，并追踪水稻叶片中的病原体。基于该组装系统，我们构建了一系列适用于经典双酶切克隆的蛋白质表达和启动子探针载体。这些载体系统能够克隆来自Xoo和Xoc菌株的所有感兴趣的基因或启动子。我们的模块化组装系统代表了一种通用且高效的基因表达分析工具包，它将加速对水稻黄单胞菌与水稻相互作用的研究。
我们开发了一种模块化质粒组装系统，用于将DNA模块从入门载体转移到高拷贝数的源自pHM1的载体中，并将其用于蛋白质检测、突变体互补以及叶片中病原体的追踪。
Xanthomonas oryzae pathovars oryzae (Xoo) and oryzicola (Xoc) infect rice, causing bacterial blight and bacterial leaf streak, respectively, which are two economically important bacterial diseases in paddy fields. The interactions of Xoo and Xoc with rice can be used as models for studying fundamental aspects of bacterial pathogenesis and host tissue specificity. However, an improved vector system for gene expression analysis is desired for Xoo and Xoc because some broad host range vectors that can replicate stably in X. oryzae pathovars are low‐copy number plasmids. To overcome this limitation, we developed a modular plasmid assembly system to transfer the functional DNA modules from the entry vectors into the pHM1‐derived backbone vectors on a high‐copy number basis. We demonstrated the feasibility of our vector system for protein detection, and quantification of virulence gene expression under laboratory conditions and in association with host rice and nonhost tobacco cells. This system also allows execution of a mutant complementation equivalent to the single‐copy chromosomal integration system and tracing of pathogens in rice leaf. Based on this assembly system, we constructed a series of protein expression and promoter‐probe vectors suitable for classical double restriction enzyme cloning. These vector systems enable cloning of all genes or promoters of interest from Xoo and Xoc strains. Our modular assembly system represents a versatile and highly efficient toolkit for gene expression analysis that will accelerate studies on interactions of X. oryzae with rice. We developed a modular plasmid assembly system to transfer DNA modules from entry vectors into high‐copy number pHM1‐derived vectors and use it for protein detection, mutant complementation, and tracing pathogens in leaves.
登录
查看更多内容
HrpD6 在调节米黄单胞菌 hrp-hrc-hpa 基因中的新调控作用。
影响因子：
3.5
作者：
Li, Yu-Rong;Zou, Hua-Song;Chen, Gong-You
通讯作者：
Chen, Gong-You
影响因子：
4.4
作者：
Figueroa-Cuilan, Wanda;Daniel, Jeremy J.;Brown, Pamela J. B.
通讯作者：
Brown, Pamela J. B.
影响因子：
3.2
作者：
Tsuge, Seiji;Nakayama, Takeshi;Kaku, Hisatoshi
通讯作者：
Kaku, Hisatoshi
影响因子：
4.2
作者：
Han, Sang-Wook;Park, Chang-Jin;Ronald, Pamela C.
通讯作者：
Ronald, Pamela C.
影响因子：
2.7
作者：
Li, Yilang;Yan, Yichao;Chen, Gongyou
通讯作者：
Chen, Gongyou