| Pseudomonas syringae Flood-inoculation Method in Arabidopsis
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Author:
Date:
2017-01-20
[Abstract] Pseudomonas syringae pv. tomato strain DC3000 (Pto DC3000), which causes bacterial speck disease of tomato, has been used as a model pathogen because of its pathogenicity on Arabidopsis thaliana. Here, we demonstrate a rapid and reliable flood-inoculation method based on young Arabidopsis seedlings grown on one-half strength MS medium. We also describe a method to evaluate the bacterial growth in Arabidopsis.
[摘要] 丁香假单胞菌 pv。 由于其在拟南芥中的致病性，已经将用于番茄细菌斑病的番茄株DC3000（ DC3000）用作模型病原体 >。 在这里，我们展示了一种基于在一半强度MS培养基上生长的幼苗拟南芥幼苗的快速可靠的接种方法。 我们还描述了一种评估拟南芥细菌生长的方法。
背景 A。 DC3000模型病理学被广泛用于研究微生物发病机制和植物先天免疫的分子机制（Ishiga等，2012和2016; Ishiga和Ichinose，2016）。 虽然已经开发了几种接种方法来研究该模型系统中的相互作用，但迄今报道的方法之一与自然界中发生的方法相似。
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| Preparation of Knockdown Transformants of Unicellular Charophycean Alga, Closterium peracerosum-strigosum-littorale Complex
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Author:
Date:
2016-05-20
[Abstract] To prepare the knockdown transformants of the Closterium peracerosum-strigosum-littorale (C. psl.) complex, particle bombardment was applied with a newly constructed vector (pSA0104) with an endogenous constitutive promoter fused to a DNA fragment corresponding to an antisense strand of a target gene. Using a hygromycin resistance gene (aph7”), hygromycin-resistant colonies were selected. After the second screening, integration of the vector into the genome was checked by PCR and the knockdown effect was evaluated by Western blotting using a specific antibody against the target protein.
[摘要] 为了制备鬼臼藓藻（Clonsterium peracerosum-strigosum-littorale）复合物的敲低转化体，用新构建的载体（pSA0104）应用粒子轰击，所述载体具有内源性 组成型启动子，其与对应于靶基因的反义链的DNA片段融合。 使用潮霉素抗性基因（ aph7"）选择潮霉素抗性菌落。 在第二次筛选后，通过PCR检查载体到基因组中的整合，并使用针对靶蛋白的特异性抗体通过蛋白质印迹评估敲低效应。
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