Cooley L, Kelley R, Spradling A
Department of Embryology, Carnegie Institution of Washington, Baltimore, MD 21210.
Science. 1988 Mar 4;239(4844):1121-8. doi: 10.1126/science.2830671.
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 element 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.
本文描述了一种通用的遗传方法，用于鉴定和克隆影响果蝇任何可识别表型的基因。构建了这样的菌株，其中单个P转座因子的插入导致了新的突变，极大地简化了对受影响基因的遗传和分子分析。诱变是通过将两个菌株杂交启动的，每个菌株都含有一个经过特殊设计的P因子。一个编码P因子转座酶的因子（启动子）有效地动员第二个非自主转座子（诱变子），其结构便于对新的插入突变进行选择和克隆。随机的诱变子转座在不再含有启动子的单个品系中被捕获，并在那里保持稳定。该方法用于构建1300个单P因子插入品系，然后筛选隐性突变。单因子插入菌株文库将使果蝇基因的结构和功能易于关联，并且在果蝇分子遗传学中应该有许多其他应用。
