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利用秀丽隐杆线虫中的易位eT1(III;V)对端粒进行定位。

Mapping a telomere using the translocation eT1(III;V) in Caenorhabditis elegans.

作者信息

Adames K A, Gawne J, Wicky C, Müller F, Rose A M

机构信息

Department of Medical Genetics, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.

出版信息

Genetics. 1998 Nov;150(3):1059-66. doi: 10.1093/genetics/150.3.1059.

Abstract

In Caenorhabditis elegans, individuals heterozygous for a reciprocal translocation produce reduced numbers of viable progeny. The proposed explanation is that the segregational pattern generates aneuploid progeny. In this article, we have examined the genotype of arrested embryonic classes. Using appropriate primers in PCR amplifications, we identified one class of arrested embryo, which could be readily recognized by its distinctive spot phenotype. The corresponding aneuploid genotype was expected to be lacking the left portion of chromosome V, from the eT1 breakpoint to the left (unc-60) end. The phenotype of the homozygotes lacking this DNA was a stage 2 embryonic arrest with a dark spot coinciding with the location in wild-type embryos of birefringent gut granules. Unlike induced events, this deletion results from meiotic segregation patterns, eliminating complexity associated with unknown material that may have been added to the end of a broken chromosome. We have used the arrested embryos, lacking chromosome V left sequences, to map a telomere probe. Unique sequences adjacent to the telomeric repeats in the clone cTel3 were missing in the arrested spot embryo. The result was confirmed by examining aneuploid segregants from a second translocation, hT1(I;V). Thus, we concluded that the telomere represented by clone cTel3 maps to the left end of chromosome V. In this analysis, we have shown that reciprocal translocations can be used to generate segregational aneuploids. These aneuploids are deleted for terminal sequences at the noncrossover ends of the C. elegans autosomes.

摘要

在秀丽隐杆线虫中,相互易位的杂合个体产生的存活后代数量减少。提出的解释是,分离模式产生非整倍体后代。在本文中,我们研究了停滞胚胎类别的基因型。在PCR扩增中使用合适的引物,我们鉴定出一类停滞胚胎,其可以通过其独特的斑点表型容易地识别。预期相应的非整倍体基因型缺少染色体V的左半部分,从eT1断点到左侧(unc-60)末端。缺乏该DNA的纯合子的表型是2期胚胎停滞,有一个黑点,与野生型胚胎中双折射肠颗粒的位置一致。与诱导事件不同,这种缺失是由减数分裂分离模式导致的,消除了与可能添加到断裂染色体末端的未知物质相关的复杂性。我们使用了缺乏染色体V左序列的停滞胚胎来定位端粒探针。克隆cTel3中与端粒重复序列相邻的独特序列在停滞的斑点胚胎中缺失。通过检查来自第二次易位hT1(I;V)的非整倍体分离子证实了该结果。因此,我们得出结论,由克隆cTel3代表的端粒定位于染色体V的左端。在该分析中,我们表明相互易位可用于产生分离非整倍体。这些非整倍体在秀丽隐杆线虫常染色体的非交叉末端的末端序列被删除。

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