一种提高水稻FISH检出率的新方法—RFLP混合标记

A New Method to Improve Detection Rate of FISH in Rice——RFLP Mix-labeled

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 1.7kb)和 5号染色体的BAC克隆44B4(137kb) ,以及 12号染色体单个RFLPRG397(约 1.5kb)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 2 7% ,大大高于单个RFLP的检出率 (7% )。其检出率虽然低于BAC克隆 44B4(6 0 % ) ,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。

Using mix-labeled 8 RFLPs (average length is about 1.7 kb) as a probe,the authors carried out fluorescence in situ hybridization in rice, and detected the single RFLP RZ397 (1.5 kb)and the BAC clone 44B4 (137 kb) simultaneously. The detection 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 method to locate genes for its simplicity and sufficiency of RFLPs linked with the functional genes. In addition, Mix-labeled RFLP groups can be used as effective markers in karotype analysis of rice and the analysis of colinearity or synteny among cereals.