海洋科学2025，Vol.49Issue(11)：97-105,9.DOI:10.11759/hykx20250917002
利用荧光原位杂交技术在海带伸展的DNA纤维上刻画45S rRNA基因的高分辨率可视图谱
High-resolution visual mapping of 45S rRNA genes by fluo-rescence in situ hybridization of the extended DNA fibers in Saccharina japonica
摘要
Abstract
The DNA fiber-fluorescence in situ hybridization(FISH)technique offers high spatial resolution.However,so far,it has not been reported in algal cytogenetics research.To address this gap,the present study optimized the nuclear lysis time and conditions for the DNA fiber extension method to obtain high-quality genomic DNA from isolated nuclei of Saccharina japonica gametophytes.Fiber-FISH was subsequently conducted on the extended DNA fibers using fluor-ophore-labeled 18S rDNA as a probe.The resulting FISH signals exhibited a linear,beads-on-a-string arrangement along the genomic DNA fibers.This pattern was consistent with that obtained using a bacterial artificial chromosome clone containing the kelp 45S rDNA sequence.Based on these extended genomic DNA fibers,a high-resolution physical map of 45S rDNA genes was constructed,achieving a resolution of≤2.09 kb,comparable to that reported in higher plants.This map provides crucial cytological evidence for accurate kelp genome assembly.In addition,it may also serve as a reference approach for assembling tandem repeat sequences and multi-copy genes in other algal genomes.关键词
海带/荧光原位杂交(FISH)/DNA纤维/18S rDNAKey words
Saccharina japonica/fluorescence in situ hybridization/DNA fiber/18S rDNA分类
生物科学引用本文复制引用
陈思奇,蔡奕彦,张程,毕燕会,周志刚..利用荧光原位杂交技术在海带伸展的DNA纤维上刻画45S rRNA基因的高分辨率可视图谱[J].海洋科学,2025,49(11):97-105,9.基金项目
国家自然科学基金(32172963 （）
41376136) National Natural Science Foundation of China,Nos.32172963,41376136 （）
