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首页> 外文期刊>Molecular and Cellular Biology >Molecular cloning of fused, a gene required for normal segmentation in the Drosophila melanogaster embryo.
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Molecular cloning of fused, a gene required for normal segmentation in the Drosophila melanogaster embryo.

机译：融合的分子克隆，这是果蝇黑腹果蝇胚胎正常分段所必需的基因。

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摘要

Using the chromosomal walk technique, we isolated recombinant lambda bacteriophage and cosmid clones spanning 250 kilobases (kb) in the 17C-D region of the X chromosome of Drosophila melanogaster. This region was known to contain the segment polarity gene fused. Several lethal fused mutations were used to define more precisely the localization of this locus. Southern analysis of genomic DNA revealed that all of them were relatively large deficiencies, the smallest one being 40 kb long. None of the 12 viable fused mutations examined possessed detectable alterations. We isolated a cosmid containing an insertion covering the entire smallest fused deletion (40 kb). We injected this DNA into fused mutant embryos and obtained a partial phenotypic rescue of the embryonic pattern, indicating that this region contained all the sequences necessary for the embryonic expression of the fu+ gene. Within this DNA, a subclone of 14 kb codes for poly(A)+ RNAs of 3.5, 2.5, 1.6, and 1.3 kb detected in embryos from various developmental stages as well as in adults. All these transcripts showed the same developmental expression. This transcribed region was injected into fused mutant embryos, and once again we obtained a partial rescue of the embryonic phenotype, confirming that this region contained at least the fused gene.
机译：使用染色体行走技术，我们分离了重组λ噬菌体和粘粒克隆，其在果蝇X染色体的X染色体的17C-D区跨越了250千个碱基（kb）。已知该区域包含融合的区段极性基因。使用几个致命的融合突变来更精确地定义该基因座的定位。对基因组DNA的Southern分析显示，它们全部都是相对较大的缺陷，最小的缺陷是40 kb长。检查的12个可行的融合突变中没有一个具有可检测的变化。我们分离出含有覆盖整个最小融合缺失（40 kb）的插入片段的粘粒。我们将此DNA注入融合的突变体胚胎中，并获得了部分胚胎型的表型拯救，表明该区域包含了fu +基因的胚胎表达所必需的所有序列。在该DNA中，14 kb的亚克隆编码了3.5，2.5，1.6和1.3 kb的poly（A）+ RNA，它们分别来自不同发育阶段的胚胎以及成年后。所有这些成绩单显示相同的发展表达。将该转录区域注入融合的突变体胚胎中，并且我们再次获得了胚胎表型的部分拯救，证实了该区域至少包含融合基因。

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