| RNA Isolation and Northern Blot Analysis
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Author:
Date:
2014-03-20
[Abstract] The northern blot is a technique used in molecular biology research to study gene expression by detection of RNA in a sample. With northern blotting it is possible to observe particular gene expression levels during differentiation, morphogenesis, as well as abnormal or diseased conditions. Here, we examine ATF3, ATF4, and GADD153 gene expression profiles by northern blot in Vero cells and H1299 cells after IBV infection. RNA was extracted in IBV (infectious bronchitis virus) infected cells and electrophoresis was used to separate the RNA sample. RNA was transferred from the electrophoresis ...
[摘要] Northern印迹是在分子生物学研究中用于通过检测样品中的RNA来研究基因表达的技术。 使用Northern印迹，可以在分化，形态发生以及异常或疾病状况期间观察到特定的基因表达水平。 在这里，我们检查ATF3，ATF4和GADD153基因表达谱通过北部污点在Vero细胞和H1299细胞后IBV感染。 在IBV（感染性支气管炎病毒）感染的细胞中提取RNA，并使用电泳分离RNA样品。 通过毛细管转移将RNA从电泳凝胶转移到印迹膜上。 使用与靶序列的一部分互补的杂交探针检测特异性mRNA。 通过RT-PCR制备探针，并使用DIG标记试剂盒通过地高辛（DIG）标记。
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| Immunofluorescence Analysis of Yeast Protein
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Author:
Date:
2012-07-05
[Abstract] Many important regulatory proteins such as transcription factors are regulated through subcellular localization. Protein localization can be examined by fusing a GFP tag. However, GFP is relatively big in size, and potentially may affect correct protein localization. Several small tags have been developed, such as myc, HA or Flag. By using immunostain and fluorescence microscopy as described in this protocol, one can easily probe the regulation of a selected yeast protein with the application of the aforementioned small tags.
[摘要] 很多重要调控蛋白如转录因子通过亚细胞定位调控。蛋白定位可以通过融和一个GFP标签来检测。尽管如此，GFP是相当的大的，可能作用于正确的蛋白定位。若干个小标签蛋白已经发展起来，如myc,HA或是Flag。通过使用以下描述的免疫染色和荧光显微镜方法，研究者们可以很容易的检测带有这些小标签酵母蛋白的调控。
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