TERACHI Toru Kyoto Sangyo University, Faculty of Engineering, Associate Professor, 工学部, 助教授 (90202192)
NAKAMURA Chiharu Kyoto Sangyo University, Faculty of Agriculture, Professor, 農学部, 教授 (10144601)
OGIHARA Yasunari Yokohama City University, Kihara Biological Research Institute, Associate Professor, 木原生物学研究所, 助教授 (40185533)
GOJOBORI Takashi National Institute of Genetics, Center for Information Biology, Professor, 生命情報研究センター, 教授 (50162136)
MURAI koji Fukui Prefectural University, Faculty of Biotechnology, Associate Professor, 生物資源学部, 助教授 (70261097)
|Budget Amount *help
¥26,100,000 (Direct Cost : ¥26,100,000)
Fiscal Year 1999 : ¥10,200,000 (Direct Cost : ¥10,200,000)
Fiscal Year 1998 : ¥15,900,000 (Direct Cost : ¥15,900,000)
Complete nucleotide sequence of chloroplast genome has been reported only in two Gramineae species, rice and corn. In this project, we succeeded determination of the complete sequence of wheat chloroplast genome as the third case. We used a common wheat (Triticum aestivum) cv. Chinese Spring as the material, because this cultivar has been used most extensively worldwide in genetic investigations. In addition, we constructed a contig clone bank of wheat chloroplast DNA. Also, PCR-primer sets were designed to amplify coding regions of all but two wheat chloroplast genes, using which their coding regions were successfully amplified. Main results are given below.
(1) Complete sequence of wheat chloroplast genome : Size of wheat chloroplast genome was 134,540 bp, that was 4.2% smaller than that of corn (140,387 bp) but almost the same as that of rice (134,525 bp). Sizes of three regions, large and small single copy regions (LSC and SSC) and inverted repeat (IR), were 80,347, 12,789 and 20,702 bp, respectively. This genome contained 109 genes, 4 (in duplication) coding for 4 rRNA species, 30 for tRNAs (corresponding to 20 amino acids), 70 for protein subunits and 5 of unidentified functions.
(2) Contig clone bank : A chloroplast DNA bank consisting of 55 contig clones (some are partially overlapping) that cover entire regions of LSC, SSC and IR was constructed. Average size of the clones was 2,246 bp.
(3) PCR primer sets for amplification of wheat chloroplast genes : A pair of PCR primers for each of 107 chloroplast genes were designed. Using these, small amounts of amplified DNAs were prepared for the coding regions of all but two wheat chloroplast genes.