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首页> 外文学位 >Isolation and characterization of a triosephosphate isomerase gene and a root-specific gene from rice, and analysis of their regulatory sequences in transgenic tobacco and rice.
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Isolation and characterization of a triosephosphate isomerase gene and a root-specific gene from rice, and analysis of their regulatory sequences in transgenic tobacco and rice.

机译：水稻中的磷酸三糖异构酶基因和根特异性基因的分离，鉴定，以及在转基因烟草和水稻中的调控序列分析。

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摘要

Genomic clones encoding cytosolic triosephosphate isomerase (TPI) and a root-specific gene, and their corresponding cDNA clones have been isolated and characterized from rice. TPI catalyzes the interconversion of dihydroxyacetone phosphate and D-glyceraldehyde-3-phosphate, an essential isomerization reaction in the glycolytic pathway. Using cytosolic TPI cDNA as a probe, it was found that the rice genome contains a single copy of the TPI gene. Northern blot analysis indicates that TPI is expressed in all vegetative tissues and rice suspension calli. Rice cytosolic TPI gene expression in roots and culms can be induced by anaerobiosis but is not light-regulated. The organization of exon-intron boundaries in rice TPI gene is identical to that of maize, and is conserved with that of TPI genes from animals. The coding sequence of the root-specific gene was found to be a member of a small gene family, which shares high sequence homology with a maize cDNA that has been shown to be specifically expressed in young maize roots. The 5{dollar}spprime{dollar} upstream sequences of both the TPI and the root-specific genes were fused to {dollar}beta{dollar}-glucuronidase (GUS) reporter gene and these chimeric constructs were stably introduced into rice (Oryza sativa L. cv Taipei 309) by protoplast electroporation and into tobacco by Agrobacterium-mediated transformation. The 5{dollar}spprime{dollar} upstream sequence of TPI gene can direct high GUS gene expression in transgenic tobacco, whereas in transgenic rice plants, no GUS activity can be detected. Assays of transient expression using microprojectile bombardment also did not give any GUS expression. It is hypothesized that an essential cis-acting element(s) of the TPI gene is required for its expression in transgenic rice. The 5{dollar}spprime{dollar} upstream sequence of the root-specific clone directed high level and cell-specific GUS expression in the roots of a transgenic rice plant. However, high GUS activity was also detected in the leaves of the same plant. These findings indicate that the regulation of root-specificity in rice may be complex and probably involves the participation of sequences other than the 5{dollar}spprime{dollar} flanking region.
机译：已经从水稻中分离并鉴定了编码胞质三糖磷酸异构酶（TPI）和根特异性基因的基因组克隆，以及它们相应的cDNA克隆。 TPI催化磷酸二羟基丙酮和D-甘油醛-3-磷酸酯的相互转化，这是糖酵解途径中必不可少的异构化反应。使用胞质TPI cDNA作为探针，发现水稻基因组包含TPI基因的单个拷贝。 Northern印迹分析表明TPI在所有营养组织和水稻悬液愈伤组织中表达。厌氧菌可以诱导水稻根和茎中的胞质TPI基因表达，但不受光调节。水稻TPI基因的外显子-内含子边界的组织与玉米相同，与动物的TPI基因的保守。发现根特异性基因的编码序列是一个小基因家族的成员，该家族与玉米cDNA具有很高的序列同源性，玉米cDNA已显示在年轻的玉米根中特异性表达。将TPI和根特异性基因的5 {sp} e {dol}上游序列与{dol}β{dol}-葡糖醛酸糖苷酶（GUS）报告基因融合，并将这些嵌合构建体稳定导入水稻（Oryza sativa） L. cv Taipei 309）通过原生质体电穿孔并通过农杆菌介导转化为烟草。 TPI基因的5 {sp} e {dollar}上游序列可以指导转基因烟草中高GUS基因的表达，而在转基因水稻植物中，无法检测到GUS活性。使用微粒轰击的瞬时表达测定也未给出任何GUS表达。假设TPI基因在转基因水稻中表达需要必需的顺式作用元件。根特异性克隆的5 {sp}}美元上游序列指导转基因水稻植株根中高水平和细胞特异性GUS表达。但是，在同一株植物的叶片中也检测到高GUS活性。这些发现表明，水稻根系特异性的调控可能很复杂，可能涉及除5 {sp} e {dollar}侧翼区域之外的序列的参与。

著录项

  • 作者

    Xu, Yong.;

  • 作者单位

    Texas A&M University.;

  • 授予单位 Texas A&M University.;
  • 学科 Biology Molecular.; Biology Botany.
  • 学位 Ph.D.
  • 年度 1993
  • 页码 133 p.
  • 总页数 133
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子遗传学;植物学;
  • 关键词

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