Plant, Animal & Microbe Genomes X Conference Plant, Animal & Microbe Genomes X ConferenceJanuary 12-16, 2002
Town & Country Convention Center
San Diego, CA
Poster: Genome Sequencing & ESTs
As the first step toward understanding of the whole genetic system of legume species, we have started a large-scale genome analysis of a model legume, Lotus japonicus. This project adopted various types of approaches for structural analysis of the genome, which include construction of genomic and cDNA libraries, collection of ESTs, serial analysis of gene expression (SAGE), genome sequencing, and generation of a genetic linkage map. For genome sequencing, we have constructed a genomic library of L. japonicus accession MG-20 in a transformation competent artificial chromosome (TAC) vector. Seed clones for sequencing were selected by high throughput PCR screening using primer sets designed on the basis of ESTs and cDNA markers of L. japonicus and other legumes. The nucleotide sequence of each selected clone was determined according to the bridging shotgun method with an approximate redundancy of 5, followed by gene assignment by similarity search and computer prediction. As of October 2001, a total of 563 seed clones have been selected. One hundred and eleven of them are in the library phase, 3 are being sequenced, 86 are in the finishing phase, 52 are being, and 50 have been annotated. Mapping of the seed clones has also been performed by generating PCR based molecular markers from sequence information, and 85 clones have been mapped on the linkage map. The sequence data and marker information are available through the web database at http://www.kazusa.or.jp/lotus.