INSERTIONAL MUTAGENESIS OF THE DROSOPHILA GENOME WITH SINGLE P-ELEMENTS
INSERTIONAL MUTAGENESIS OF THE DROSOPHILA GENOME WITH SINGLE P-ELEMENTS
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DOI：
10.1126/science.2830671
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发表时间：
1988-03-04
期刊：
影响因子：
56.9
通讯作者：
SPRADLING, A
中科院分区：
文献类型：
--
作者：
COOLEY, L;KELLEY, R;SPRADLING, A
描述了一种通用的遗传方法，用于识别和克隆影响任何可识别表型的果蝇黑腹果蝇基因。构建的菌株中，单个P转座元件的插入导致了新的突变，极大地简化了受影响基因的遗传和分子分析。突变是通过使两个菌株杂交来启动的，每个菌株都含有一个特殊设计的P元素。其中一个元件(Jumpstarter)编码P元件转座酶，它有效地动员第二个非自主转座子(Mutator)，其结构便于选择和克隆新的插入突变。随机突变因子换位被捕获在不再包含Jumpstarter的个股中，在那里它们保持稳定。用这种方法构建了1300个单P元素插入群体，并对这些群体进行了隐性突变的筛选。单元件插入菌株文库将使果蝇基因的结构和功能易于关联，并将在果蝇分子遗传学中有许多其他应用。
A versatile genetic method for identifying and cloning Drosophila melanogaster genes affecting any recognizable phenotype is described. Strains are constructed in which the insertion of a single P transposable elements has caused a new mutation, greatly simplifying the genetic and molecular analysis of the affected gene. Mutagenesis is initiated by crossing two strains, each of which contains a specially designed P element. One element (jumpstarter), encoding P element transposase, efficiently mobilizes the second nonautonomous transposon (mutator), whose structure facilitates selection and cloning of new insertion mutations. Random mutator transpositions are captured in individual stocks that no longer contain jumpstarter, where they remain stable. This method was used to construct 1300 single P element insertion stocks which were then screened for recessive mutations. A library of single-element insertion strains will allow the structure and function of Drosophila genes to be readily correlated, and should have many other applications in Drosophila molecular genetics.