| 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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| Gentiobiose Feeding in Gentian in vitro Overwintering Buds or Plantlets
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Author:
Date:
2015-06-20
[Abstract] To study the functions of sugars in plants, feeding experiment is one of the most common and easy methods. However, the traditional method, e.g., a floating of leaf discs on sugar-containing solution seems to have an insufficient efficiency of sugar uptake, despite of high osmotic and injury effects. This is a protocol to feed oligosaccharide gentiobiose into in vitro cultured tissues of gentian. This protocol enables to incorporate gentiobiose into intact tissues without exposure to osmotic stress and may be useful to other plant species that are able to propagate by shoot tip culture.
[摘要] 为了研究植物中糖的功能，饲养实验是最常见和最容易的方法之一。 然而，传统方法，例如，在含糖溶液上漂浮叶盘似乎具有不充分的糖吸收效率，尽管具有高的渗透和损伤效应。 这是一种将寡糖龙胆二糖供给到体外培养的龙胆的组织中的方案。 该方案能够将龙胆二糖掺入完整组织而不暴露于渗透胁迫，并且可以用于能够通过芽尖培养繁殖的其它植物物种。
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