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利用酵母位点特异性重组酶在果蝇中产生雌性生殖系嵌合体。

Use of a yeast site-specific recombinase to produce female germline chimeras in Drosophila.

作者信息

Chou T B, Perrimon N

机构信息

Howard Hughes Medical Institute, Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115.

出版信息

Genetics. 1992 Jul;131(3):643-53. doi: 10.1093/genetics/131.3.643.

Abstract

We describe an efficient method for generating female germline mosaics by inducing site-specific homologous mitotic recombination with a yeast recombinase (FLP) which is driven by a heat shock promoter. These germline mosaics are produced in flies heterozygous for the agametic, germline-dependent, dominant female sterile (DFS) mutation ovoD1, where only flies possessing germline clones are able to lay eggs. This method, the "FLP-DFS" technique, is very efficient because more than 90% of females with germline clones can be recovered. We show that this heat-inducible, site-specific mitotic recombination system does not affect viability and that the germline clones recovered are physiologically the same as those created by X-ray induced mitotic recombination. We describe the parameters of FLP-recombinase induced germline mitotic recombination and the use of the "FLP-DFS" technique to analyze the maternal effect of X-linked zygotic lethal mutations.

摘要

我们描述了一种通过热休克启动子驱动的酵母重组酶(FLP)诱导位点特异性同源有丝分裂重组来产生雌性种系嵌合体的有效方法。这些种系嵌合体是在无配子、种系依赖性、显性雌性不育(DFS)突变ovoD1杂合的果蝇中产生的,其中只有拥有种系克隆的果蝇才能产卵。这种方法,即“FLP-DFS”技术,非常有效,因为超过90%具有种系克隆的雌性能够被回收。我们表明,这种热诱导的位点特异性有丝分裂重组系统不会影响生存能力,并且回收的种系克隆在生理上与通过X射线诱导的有丝分裂重组产生的种系克隆相同。我们描述了FLP重组酶诱导种系有丝分裂重组的参数,以及使用“FLP-DFS”技术分析X连锁合子致死突变的母体效应。

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