| 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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| Transformation of the Cyanobacterium Leptolyngbya boryana by Electroporation
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Author:
Date:
2015-12-20
[Abstract] Leptolyngbya boryana (L. boryana) (formerly Plectonema boryanum) is a versatile, filamentous cyanobacterium that has the ability to fix nitrogen under microoxic conditions and to grow heterotrophically with glucose in the dark, providing an excellent system to investigate photosynthesis, nitrogen fixation, and their regulatory mechanisms. While L. boryana is not naturally transformable different from the unicellular cyanobacterium Synechocystis sp. PCC 6803, it can be transformed by electroporation. Here we describe the transformation of L. boryana by electroporation to isolate mutants in which a targeted gene is disrupted.
[摘要] Leptolyngbya boryana （ L。boryana ）（以前称为Plectonema boryanum）是一种多用途的丝状蓝细菌，具有在微氧条件下固氮的能力， 在黑暗中与葡萄糖异养生长，提供了研究光合作用，固氮和它们的调节机制的优良系统。 而 L。 boryana 不是天然可转化的，不同于单细胞蓝细菌集胞藻。 PCC 6803，它可以通过电穿孔转化。 这里我们描述了 L的变换。 boryana通过电穿孔分离其中靶基因被破坏的突变体。
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