Kato Tomohiko, Sato Shusei, Nakamura Yasukazu, Kaneko Takakazu, Asamizu Erika, Tabata Satoshi
Kazusa DNA Research Institute, 2-6-7 Kazusa-kamatari, Kisarazu, Chiba 292-0818, Japan.
DNA Res. 2003 Dec 31;10(6):277-85. doi: 10.1093/dnares/10.6.277.
We determined the nucleotide sequences of 64 TAC (transformation-competent artificial chromosome) clones selected from genomic libraries of Lotus japonicus accession Miyakojima MG-20 based on the sequence information of expressed sequence tags (ESTs), cDNAs, genes and DNA markers from L. japonicus and other legumes. The length of the DNA regions sequenced in this study was 6,370,255 bp, and the total length of the L. japonicus genome sequenced so far is 32,537,698 bp together with the nucleotide sequences of 256 TAC clones previously reported. Five hundred forty-eight potential protein-encoding genes with known or predicted functions, 127 gene segments and 224 pseudogenes were assigned to the newly sequenced regions by computer prediction and similarity searches against the sequences in protein and EST databases. Based on the nucleotide sequences of the clones, simple sequence repeat length polymorphism (SSLP) or derived cleaved amplified polymorphic sequence (dCAPS) markers were generated, and each clone was genetically localized onto the linkage map of two accessions of L. japonicus, MG-20 and Gifu B-129. The sequence data, gene information and mapping information are available through the World Wide Web at http://www.kazusa.or.jp/lotus/.
我们根据来自日本百脉根(Lotus japonicus)和其他豆科植物的表达序列标签(EST)、cDNA、基因及DNA标记的序列信息,从日本百脉根宫古岛MG - 20的基因组文库中挑选了64个转化 competent 人工染色体(TAC)克隆,并测定了它们的核苷酸序列。本研究中测序的DNA区域长度为6,370,255 bp,到目前为止,已测序的日本百脉根基因组总长度为32,537,698 bp,这还包括之前报道的256个TAC克隆的核苷酸序列。通过计算机预测以及与蛋白质和EST数据库中的序列进行相似性搜索,在新测序区域中确定了548个具有已知或预测功能的潜在蛋白质编码基因、127个基因片段和224个假基因。基于这些克隆的核苷酸序列,生成了简单序列重复长度多态性(SSLP)或衍生酶切扩增多态性序列(dCAPS)标记,并将每个克隆定位到日本百脉根两个品种MG - 20和岐阜B - 129的连锁图谱上。序列数据、基因信息和定位信息可通过万维网在http://www.kazusa.or.jp/lotus/获取。