一种提高水稻FISH检出率的新方法-RFLP混合标记

摘要

Using mix-labeled 8 RFLPs (average length is about 1.7 kb) as a probe，the autho rs carried out fluorescence in situ hybridization in rice, and detected the sing le RFLP RZ397 (1.5 kb)and the BAC clone 44B4 (137 kb) simultaneously. The detec tion rate of mix-labeled RFLP was 27%, much higher than single-labeled RFLP (7%) , though lower than BAC clone (60%). Mix-labeled RFLP is an easy and effective m ethod to locate genes for its simplicity and sufficiency of RFLPs linked with th e functional genes. In addition, Mix-labeled RFLP groups can be used as effectiv e markers in karotype analysis of rice and the analysis of colinearity or synten y among cereals.%分别以水稻1号染色体上混合标记的8个紧密连锁的RFLP(平均约1.7 kb)和5号染色体的BAC克隆44B4 (137 kb)，以及12号染色体单个RFLP RG397（约1.5 kb）为探针,在水稻染色体上进行了荧光原位杂交(FISH)。结果表明，RFLP混合标记杂交的检出率为27%，大大高于单个RFLP的检出率(7%)。其检出率虽然低于BAC克隆44B4(60%)，但它具有程序简单易行的特点，使基因原位定位更加高效。由于水稻中与已知功能基因紧密连锁的RFLP标记具有数量丰富、分布密集等优势，揭示了混合标记的RFLP在禾本科植物同线性和共线性分析中的广阔应用前景。此外，混合标记的RFLP还可以用于染色体的准确识别和核型分析。