Development of Gateway Binary Vector Series with Four Different Selection Markers for the Liverwort Marchantia polymorpha.
Development of Gateway Binary Vector Series with Four Different Selection Markers for the Liverwort Marchantia polymorpha.
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DOI：
10.1371/journal.pone.0138876
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Kohchi T
中科院分区：
文献类型：
--
作者：
Ishizaki K;Nishihama R;Ueda M;Inoue K;Ishida S;Nishimura Y;Shikanai T;Kohchi T
我们以前报道了农杆菌介导的转化方法地钱使用潮霉素磷酸转移酶基因作为标记与潮霉素的选择。在这项研究中，我们开发了三个额外的标记M。多形转化：用庆大霉素选择的庆大霉素3 ′-乙酰转移酶基因;用氯磺隆选择的突变乙酰乳酸合酶基因;和用G418选择的新霉素磷酸转移酶II基因。基于这四个标记基因，我们构建了一系列Gateway双元载体，用于转基因实验。多形。来自花椰菜花叶病毒的35 S启动子和用于组成型和热诱导型表达的内源启动子用于创建这些载体。使用的报告基因和标签为Citrine、3×Citrine、Citrine-NLS、TagRFP、tdTomato、tdTomato-NLS、GR、SRDX、SRDX-GR、GUS、ELuc（PEST）和3×FLAG。这些载体命名为pMpGWB系列，将有助于新兴模式植物M的分子遗传分析。多形。
We previously reported Agrobacterium-mediated transformation methods for the liverwort Marchantia polymorpha using the hygromycin phosphotransferase gene as a marker for selection with hygromycin. In this study, we developed three additional markers for M. polymorpha transformation: the gentamicin 3'-acetyltransferase gene for selection with gentamicin; a mutated acetolactate synthase gene for selection with chlorsulfuron; and the neomycin phosphotransferase II gene for selection with G418. Based on these four marker genes, we have constructed a series of Gateway binary vectors designed for transgenic experiments on M. polymorpha. The 35S promoter from cauliflower mosaic virus and endogenous promoters for constitutive and heat-inducible expression were used to create these vectors. The reporters and tags used were Citrine, 3×Citrine, Citrine-NLS, TagRFP, tdTomato, tdTomato-NLS, GR, SRDX, SRDX-GR, GUS, ELuc(PEST), and 3×FLAG. These vectors, designated as the pMpGWB series, will facilitate molecular genetic analyses of the emerging model plant M. polymorpha.