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用于荧光原位杂交(FISH)分析的寡核苷酸替代重复序列pAs1、pSc119.2、pTa - 535、pTa71、CCS1和pAWRC.1的作用。

Oligonucleotides replacing the roles of repetitive sequences pAs1, pSc119.2, pTa-535, pTa71, CCS1, and pAWRC.1 for FISH analysis.

作者信息

Tang Zongxiang, Yang Zujun, Fu Shulan

机构信息

State Key Laboratory of Plant Breeding and Genetics, Sichuan Agricultural University, Wenjiang, Chengdu, 611130, Sichuan, People's Republic of China.

出版信息

J Appl Genet. 2014 Aug;55(3):313-8. doi: 10.1007/s13353-014-0215-z. Epub 2014 Apr 30.

Abstract

Hybrids derived from wheat (Triticum aestivum L.) × rye (Secale cereale L.) have been widely studied because of their important roles in wheat cultivar improvement. Repetitive sequences pAs1, pSc119.2, pTa-535, pTa71, CCS1, and pAWRC.1 are usually used as probes in fluorescence in situ hybridization (FISH) analysis of wheat, rye, and hybrids derived from wheat × rye. Usually, some of these repetitive sequences for FISH analysis were needed to be amplified from a bacterial plasmid, extracted from bacterial cells, and labeled by nick translation. Therefore, the conventional procedure of probe preparation using these repetitive sequences is time-consuming and labor-intensive. In this study, some appropriate oligonucleotide probes have been developed which can replace the roles of repetitive sequences pAs1, pSc119.2, pTa-535, pTa71, CCS1, and pAWRC.1 in FISH analysis of wheat, rye, and hybrids derived from wheat × rye. These oligonucleotides can be synthesized easily and cheaply. Therefore, FISH analysis of wheat and hybrids derived from wheat × rye using these oligonucleotide probes becomes easier and more economical.

摘要

源自小麦(Triticum aestivum L.)×黑麦(Secale cereale L.)的杂种因其在小麦品种改良中的重要作用而受到广泛研究。重复序列pAs1、pSc119.2、pTa - 535、pTa71、CCS1和pAWRC.1通常用作小麦、黑麦以及源自小麦×黑麦的杂种的荧光原位杂交(FISH)分析中的探针。通常,这些用于FISH分析的重复序列中的一些需要从细菌质粒中扩增、从细菌细胞中提取并通过缺口平移进行标记。因此,使用这些重复序列制备探针的传统方法既耗时又费力。在本研究中,已经开发出了一些合适的寡核苷酸探针,它们可以在小麦、黑麦以及源自小麦×黑麦的杂种的FISH分析中取代重复序列pAs1、pSc119.2、pTa - 535、pTa71、CCS1和pAWRC.1的作用。这些寡核苷酸可以轻松且廉价地合成。因此,使用这些寡核苷酸探针进行小麦以及源自小麦×黑麦的杂种的FISH分析变得更加容易且经济。

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