Chemical mutagens, transposons, and transgenes to interrogate gene function in Drosophila melanogaster.
Chemical mutagens, transposons, and transgenes to interrogate gene function in Drosophila melanogaster.
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DOI：
10.1016/j.ymeth.2014.02.025
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发表时间：
2014-06-15
期刊：
影响因子：
4.8
通讯作者：
Bellen, Hugo J.
中科院分区：
文献类型：
--
作者：
Venken, Koen J. T.;Bellen, Hugo J.
关键词：
遗传学、基因和染色体遗传的研究是由托马斯摩根在年首次在果蝇中发现可见的突变时发起的。该领域扩大了赫尔曼穆勒在1926年开始的工作，当时他使用X射线开发了第一个平衡染色体。今天，平衡器在保持突变和转基因方面仍然是非常宝贵的，但工具库已经大大扩展，并且已经开发了许多新方法，其中许多依赖于基因组序列和转座因子的可用性。基于化学诱变或转座因子的正向遗传筛选已经导致了通过特定表型测定探测的过程中所涉及的许多新参与者的无偏鉴定。反向遗传方法依赖于在整个基因组中分布的一组精心选择的转座子插入的可用性，以允许操纵每个插入附近的区域。最后，使用转座子用单拷贝转基因转化果蝇的能力或使用ΦC31整合酶进行位点特异性整合的能力已经允许许多操作，包括创建和整合基因组拯救构建体、产生重复、RNAi敲除技术、二元表达系统如GAL 4/UAS系统以及其他方法的能力。在这里，我们将讨论最有用的方法来询问果蝇基因组在体内集中在化学诱变，转座子和转基因。基于核酸酶和RNAi技术的基因组工程方法在以下章节中讨论。
The study of genetics, genes, and chromosomal inheritance was initiated by Thomas Morgan in when the first visible mutations were identified in fruit flies. The field expanded upon the work initiated by Herman Muller in 1926 when he used X-rays to develop the first balancer chromosomes. Today, balancers are still invaluable to maintain mutations and transgenes but the arsenal of tools has expanded vastly and numerous new methods have been developed, many relying on the availability of the genome sequence and transposable elements. Forward genetic screens based on chemical mutagenesis or transposable elements have resulted in the unbiased identification of many novel players involved in processes probed by specific phenotypic assays. Reverse genetic approaches have relied on the availability of a carefully selected set of transposon insertions spread throughout the genome to allow the manipulation of the region in the vicinity of each insertion. Lastly, the ability to transform Drosophila with single copy transgenes using transposons or site-specific integration using the ΦC31 integrase has allowed numerous manipulations, including the ability to create and integrate genomic rescue constructs, generate duplications, RNAi knock-out technology, binary expression systems like the GAL4/UAS system as well as other methods. Here, we will discuss the most useful methodologies to interrogate the fruit fly genome in vivo focusing on chemical mutagenesis, transposons and transgenes. Genome engineering approaches based on nucleases and RNAi technology are discussed in following chapters.
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影响因子：
4.8
作者：
Beumer, Kelly J.;Carroll, Dana
通讯作者：
Carroll, Dana
DOI：
10.1534/g3.112.003681
发表时间：
2012-10
期刊：
G3 (Bethesda, Md.)
影响因子：
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作者：
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通讯作者：
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影响因子：
3.3
作者：
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通讯作者：
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