Rice Genome Research Program, National Institute of Agrobiolog-cal Resources/Institute of Society for Techno-Innovation of Agriculture, Forestry and Fisheries, 1-2 Kannondai 2-chome, Tsukuba, 305, Ibaraki, Japan.
Theor Appl Genet. 1994 Nov;89(6):728-34. doi: 10.1007/BF00223712.
Generating sequence-tagged sites (STSs) is a prerequisite to convert a genetic map to a physical map. With the help of sequence information from these STSs one can also isolate specific genes. For these purposes, we have designed PCR primer sets, of 20 bases each, by reference to sequences of restriction fragment length polymorphism (RFLP) landmarkers consisting of rice genomic clones. These markers were evenly distributed over the 12 chromosomes and were shown to be single copy by Southern-blot analysis. With improved PCR protocols, 63 standard STS landmarkers in the rice genome were generated. Similarity searches of all partial sequences of RFLP landmarkers by the FASTA algorithm showed that 2 of the 63 RFLP landmarkers, G357 and G385, contained part of the ORFs of aspartate aminotransferase and protein kinase, respectively.
生成序列标记位点（STS）是将遗传图谱转化为物理图谱的前提条件。借助这些 STS 的序列信息，还可以分离特定的基因。为此，我们参考了由水稻基因组克隆组成的限制片段长度多态性（RFLP）标记物的序列，设计了 20 个碱基的 PCR 引物对。这些标记物均匀分布在 12 条染色体上，通过 Southern 印迹分析显示为单拷贝。通过改进的 PCR 方案，在水稻基因组中生成了 63 个标准 STS 标记物。通过 FASTA 算法对所有 RFLP 标记物的部分序列进行相似性搜索表明，63 个 RFLP 标记物中的 2 个，G357 和 G385，分别包含天冬氨酸转氨酶和蛋白激酶的部分 ORF。
