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通过多色荧光原位杂交对甜菜染色体上的酵母人工染色体(YACs)和单拷贝基因Hs1(pro-1)进行高分辨率定位。

High-resolution mapping of YACs and the single-copy gene Hs1(pro-1) on Beta vulgaris chromosomes by multi-colour fluorescence in situ hybridization.

作者信息

Desel C, Jung C, Cai D, Kleine M, Schmidt T

机构信息

Institute of Crop Science and Plant Breeding, Christian Albrecht University of Kiel, Germany.

出版信息

Plant Mol Biol. 2001 Jan;45(1):113-22. doi: 10.1023/a:1006405911442.

Abstract

Fluorescence in situ hybridization (FISH) is a powerful approach for physical mapping of DNA sequences along plant chromosomes. Nematode-resistant sugar beets (Beta vulgaris) carrying a Beta procumbens translocation were investigated by FISH with two differentially labelled YACs originating from the translocation. At mitotic metaphases, the translocation was identified with both YACs in the terminal region on a pair of chromosomes. Meiotic chromosomes, representing a far more extended hybridization target, were used to determine the orientation of YACs with respect to chromosomal domains in combination with chromosomal landmark probes for telomeres and centromeres. The in situ detection of plant single-copy sequences is technically difficult, and the wild beet translocation was used to explore the potential resolution of the FISH approach and to introduce the chromosomal mapping of single-copy genes into genome analysis of Beta species. An internal fragment of the nematode resistance gene Hs1(pro-1), 684 bp long, was detected on both chromatids of different Beta chromosomes and represents one of the shortest unique DNA sequences localized on mitotic plant chromosomes so far. Comparative chromosomal mapping of the 684 bp Hs1(pro-1) probe in the translocation line, a monosomic addition line and in B. procumbens revealed the origin of the wild beet translocation leading to nematode-resistant sugar beets.

摘要

荧光原位杂交(FISH)是一种用于沿植物染色体对DNA序列进行物理定位的强大方法。利用来自易位的两个差异标记的酵母人工染色体(YAC),通过FISH对携带平卧甜菜易位的抗线虫甜菜(Beta vulgaris)进行了研究。在有丝分裂中期,在一对染色体的末端区域用两个YAC都鉴定出了易位。减数分裂染色体代表了一个更为扩展的杂交靶标,结合用于端粒和着丝粒的染色体标记探针,用于确定YAC相对于染色体结构域的方向。植物单拷贝序列的原位检测在技术上具有挑战性,利用野生甜菜易位来探索FISH方法的潜在分辨率,并将单拷贝基因的染色体定位引入甜菜属物种的基因组分析中。在不同甜菜染色体的两条染色单体上都检测到了684 bp长的抗线虫基因Hs1(pro-1)的内部片段,它是迄今为止定位在植物有丝分裂染色体上的最短的独特DNA序列之一。对易位系、单体附加系和平卧甜菜中684 bp Hs1(pro-1)探针的比较染色体定位揭示了导致抗线虫甜菜的野生甜菜易位的起源。

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