Generation of 919 expressed sequence tags from immature flower buds and gene expression analysis using expressed sequence tags in the model plant Lotus japonicus

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概要

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• Lotus japonicus has received increased attention as a potential model legume plant. In order to study gene expression in reproductive organs and to identify genes that play a crucial function in sexual reproduction, we constructed a cDNA library from immature flower buds containing anthers at the stage of developing tapetum cells in L. japonicus, and characterized 919 expressed sequence tags (ESTs) randomly selected from a cDNA library of the immature flower buds. The 919 ESTs analyzed were clustered into 821 non-redundant EST groups. As a result of a database search, 436 groups (53%) out of the 821 groups showed sequence similarity to genes registered in the public database. Out of these 436 groups, 109 groups showed similarity to genes encoding hypothetical proteins whose function had not yet been estimated. Three hundred eighty five groups (47%) showed no significant homology to known sequences and were classified as novel sequences. A comparison of 821 non-redundant EST sequences and EST sequences derived from the whole plant L. japonicus revealed that 474 EST sequences derived from immature flower buds were not found in the EST sequences of the whole plant. In order to confirm the expression pattern of potential reproductive-organ specific EST clones, nine clones, which were not matched to ESTs derived from the whole plant, were selected, and RT-PCR analysis was performed on these clones. As a result of RT-PCR, we found two novel anther specific clones. One clone was homologous to a gene encoding human cleft lip and palate associated transmembrane protein (CLPTM1) like protein, and the other clone did not show a significant similarity to any genes deposited in the public database. These results indicate that ESTs analyzed here represent a valuable resource for finding reproductive-organ specific genes in Lotus japonicus.

• 日本遺伝学会の論文
• 2002-08-25

著者

• TAKAHATA Yoshihito

  Faculty of Agriculture, Iwate University

• Watanabe M

  Lab. Of Plant Reproductive Genetics Graduate School Of Life Sciences Tohoku Univ.

• 渡辺 正勝

  岡崎国立共同研究機構基礎生物学研究所

• TABATA Satoshi

  Kazusa DNA Research Institute

• Mizuho Hidetoshi

  Faculty Of Agriculture Iwate University

• Endo Makoto

  Lab. Of Biotechnology National Inst. Of Crop Sci.

• Tabata Satoshi

  Kazusa Dna Res.

• HIGASHITANI Atsushi

  Graduate School of Life Sciences, Tohoku University

• TSUCHIYA Tohru

  Faculty of Bioresources, Mie University

• MIZUNO Hidetoshi

  Graduate School of Life Sciences, Tohoku University

• Endo Makoto

  Faculty Of Agriculture Miyazaki University

• MASUKO Hiromi

  Faculty of Agriculture, Iwate University

• HAKOZAKI Hirokazu

  Faculty of Agriculture, Iwate University

• WATANABE Masao

  Faculty of Agriculture, Iwate University

• KOKUBUN Takao

  Faculty of Agriculture, Iwate University

• HAKOZAKI Hirokazu

  Graduate School of Life Sciences, Tohoku University

• Takahata Yoshihito

  Laboratory Of Plant Reproductive Genetics Graduate School Of Life Sciences Tohoku University

• Takada Yoshinobu

  Faculty Of Agriculture Iwate University

• Takahata Yoshihito

  Faculty Of Agnculture Lwate Untversity

• Higashitani A

  Kagoshima University Graduate School Of Medical And Dental Sciences Space Environmental Medicine

• Kokubun Takao

  Faculty Of Agriculture Iwate University

• Masuko Hiromi

  Faculty Of Agriculture Iwate University

• Tsuchiya T

  Life Science Research Center Mie University

• Tsuchiya Tohru

  Faculty Of Agriculture Tohoku University

• Higashitani Atsushi

  Graduate School Of Life Sciences Tohoku Univ.

• Masuko Hiromi

  Graduate School Of Life Sciences Tohoku University

• Endo M

  Lab. Of Biotechnology National Inst. Of Crop Sci.

• Tabata Satoshi

  Kazusa Dna Institute

• Watanabe Masao

  Faculty Of Agriculture Iwate University

• Higashitani Atsushi

  Kagoshima University Graduate School Of Medical And Dental Sciences Space Environmental Medicine

• Hakozaki Hirokazu

  Graduate School Of Life Sciences Tohoku University

• ENDO Makoto

  Faculty of Liberal Arts, Dokkyo University

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