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首页> 外文期刊>Genes & Genetic Systems >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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Generation of 919 expressed sequence tags from immature flower buds and gene expression analysis using expressed sequence tags in the model plant Lotus japonicus

机译：从未成熟花芽中产生919个表达序列标签并在模型植物Lotus japonicus中使用表达序列标签进行基因表达分析

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摘要

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.
机译：作为一种潜在的豆科植物典范，日本莲花已经受到越来越多的关注。为了研究生殖器官中的基因表达并鉴定在有性生殖中起关键作用的基因，我们从日本花车绒毡层细胞发育阶段的花药中构建了一个包含花药的未成熟花蕾的cDNA文库，并鉴定了919个表达序列从未成熟花蕾的cDNA文库中随机选择的标签（EST）。分析的919个EST分为821个非冗余EST组。数据库搜索的结果是，821个组中的436个组（53％）显示出与在公共数据库中注册的基因的序列相似性。在这436个组中，有109个组显示出与编码其功能尚未估计的假设蛋白质的基因相似。 385个组（47％）与已知序列没有显着同源性，被归类为新序列。比较了821个非冗余的EST序列和来源于全株日本刺槐的EST序列，发现在整个植物的EST序列中未发现来自未成熟花芽的474个EST序列。为了确认潜在的生殖器官特异性EST克隆的表达模式，选择了与来自整个植物的EST不匹配的9个克隆，并对这些克隆进行RT-PCR分析。作为RT-PCR的结果，我们发现了两个新颖的花药特异性克隆。一个克隆与编码人唇left裂相关跨膜蛋白（CLPTM1）样蛋白的基因同源，另一个克隆与存放在公共数据库中的任何基因均未显示出显着相似性。这些结果表明，本文分析的EST代表了在日本莲花中发现生殖器官特异性基因的宝贵资源。

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