棉花Oligo-FISH技术建立及其初步应用

摘要

[Objective] Fluorescence in situ hybridization (FISH) can achieve the accurate chromosomal localization of DNA sequences,which is one of the important techniques for genome in-depth study.The acquisition of chromosome-specific probes is the key for FISH application.The aim of this study is to explore a system of cotton oligonucleotides (oligos) fluorescence in situ hybridization (Oligo-FISH) technology.[Method] Based on the published cotton genome sequence,the chromosome-specific oligos library was obtained by bioinformatics method.Then,the obtained oligos were labeled through emulsion polymerase chain reaction,and used for FISH of cotton mitotic chromosome.[Results] A system of cotton Oligo-FISH technology was set up and successfully applied in cotton chromosome identification.[Conclusion] This system can be used for dissection of cotton genomes.%[目的]荧光原位杂交技术可以实现DNA序列直观准确的染色体定位,是基因组深入研究的重要技术之一.染色体特异探针的获得是该技术应用的关键.本研究旨在建立棉花寡核苷酸荧光原位杂交技术.[方法]利用已经公布棉花基因组序列数据,采用生物信息学方法获得染色体特异的寡核苷酸库,随后用乳化聚合酶链式反应方法标记成荧光探针,在棉花有丝分裂中期染色体上进行原位杂交.[结果]建立了一套棉花寡核苷酸荧光原位杂交技术体系.[结论]该体系可用于棉花单染色体识别鉴定.