FROM GENE TO PHENOTYPE IN DROSOPHILA AND OTHER ORGANISMS
FROM GENE TO PHENOTYPE IN DROSOPHILA AND OTHER ORGANISMS
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DOI：
10.1002/bies.950120609
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发表时间：
1990-06-01
期刊：
影响因子：
4
通讯作者：
KAISER, K
中科院分区：
文献类型：
--
作者：
KAISER, K
越来越多的克隆真核生物基因缺乏明确的或证明的生物学功能，提出了一个重大的挑战，在“反向遗传学”。本文描述了一种方法，该方法允许有效筛选对应于感兴趣的克隆DNA序列的基因中或附近的新病变。该技术涉及转座子诱变，然后筛选从诱变个体群体（或其后代）分离的DNA，以证明该群体含有至少一个在靶基因座处发生转座子插入的个体。通过使用聚合酶链反应（PCR），可以方便地检测人群中的罕见个体。一旦被识别，特定的个体（或其后代）通过同胞选择过程从群体中分离出来。在转座子的插入发生在靶基因座附近但不在靶基因座内的情况下，涉及转座子的不精确切除的次级事件仍然允许分离突变个体。虽然该方法是专门为果蝇的转座子诱变而开发的，但扩展到其他生物和其他诱变策略是可行的，并讨论了一些可能性。
The growing number of cloned eukaryotic genes lacking a defined or proven biological function poses a major challenge in ‘reverse genetics’. A method is described here that permits efficient screening for new lesions in, or close to, genes corresponding to cloned DNA sequences of interest. The technique involves transposon mutagenesis, followed by screening of DNA isolated from a population of mutagenised individuals (or their progeny) for evidence that the population contains at least one individual in which transposon insertion has occurred at the target locus. Detection of rare individuals within the population is facilitated by the use of the polymerase chain reaction (PCR). Once recognised, specific individuals (or their progeny) are isolated from the population by a process of sib‐selection. In cases where insertion of the transposon has occurred close to, but not within, the target locus, secondary events involving imprecise excision of the transposon will nonetheless allow the isolation of mutant individuals. Though the method was developed specifically for the transposon‐mutagenesis ofDrosophila, extensions to other organisms and to other mutagenic strategies are feasible and some of the possibilities are discussed.