Preparation of Drosophila polytene chromosome squashes for antibody labeling.
Preparation of Drosophila polytene chromosome squashes for antibody labeling.
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DOI：
10.3791/1748
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发表时间：
2010-02-09
期刊：
影响因子：
--
通讯作者：
Johansen, Kristen M
中科院分区：
文献类型：
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作者：
Cai, Weili;Jin, Ye;Johansen, Kristen M
由于果蝇三龄幼虫巨大的唾腺多线染色体所提供的细胞学优势，果蝇一直是研究染色质结构与基因调控关系的理想模式系统。在这种组织中，染色体在没有细胞分裂的情况下进行多轮复制，产生约1000个拷贝。DNA在每个复制周期后保持对齐，导致染色体大大扩大，这提供了将染色质形态与特定蛋白质定位相关联的独特机会。因此，人们对定义存在于不同基因和转录过程不同阶段的表观遗传修饰非常感兴趣。这种研究的一个重要工具是用针对酶、转录因子或组蛋白修饰的抗体标记多线染色体。这个视频协议说明了在约翰森实验室使用的挤压技术，以准备果蝇多线染色体的抗体标记。
Drosophila has long been a favorite model system for studying the relationship between chromatin structure and gene regulation due to the cytological advantages provided by the giant salivary gland polytene chromosomes of third instar larvae. In this tissue the chromosomes undergo many rounds of replication in the absence of cell division giving rise to approximately 1000 copies. The DNA remains aligned after each replicative cycle resulting in greatly enlarged chromosomes that provide a unique opportunity to correlate chromatin morphology with the localization of specific proteins. Consequently, there has been a high level of interest in defining the epigenetic modifications present at different genes and at different stages of the transcription process. An important tool for such studies is the labeling of polytene chromosomes with antibodies to the enzyme, transcription factor, or histone modification of interest. This video protocol illustrates the squash technique used in the Johansen laboratory to prepare Drosophila polytene chromosomes for antibody labeling.