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首页> 外文学位 >Cloning and in vivo analysis of a nuclear-encoded chloroplast-localized 28 kDa RNA-binding protein.
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Cloning and in vivo analysis of a nuclear-encoded chloroplast-localized 28 kDa RNA-binding protein.

机译：核编码叶绿体定位的28 kDa RNA结合蛋白的克隆和体内分析。

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摘要

A cDNA encoding a chloroplast-localized 28 kDa RNP, At28RNP, was isolated from Arabidopsis. Chloroplast import experiments and RNA UV-crosslinking have confirmed the localization and RNA-binding ability of this protein suggested by its deduced sequence. Expression analysis of At28RNP revealed that the mRNA and the encoded plastid-localized protein accumulate in the active tissues. The At28RNP protein has high sequence identity (60.6%) to a spinach chloroplast protein, So28RNP, which binds to the 3′ ends of plastid RNAs that contain transcript stabilizing inverted repeat sequences (IRs) in their untranslated regions. In vitro RNA-processing assays have shown that the So28RNP is necessary for the proper processing of a precursor RNA to a mature form. The identification and isolation of a homologous protein n Arabidopsis has allowed for the in vivo analysis of plastid localized RNP function using a reverse genetic approach. Transgenic Arabidopsis plants were generated with both increased and decreased levels of At28RNP and while protein levels were dramatically effected in both types of transgenic plants, no significant differences were found between transgenic plants and controls with respect to affects on plastid RNA maturation or accumulation. However, the stable inheritance of altered At28RNP protein expression within the transgenic lines may prove useful in further studies of RNPs in chloroplast gene expression.
机译：从 Arabidopsis 中分离出一个编码叶绿体定位的28kDa RNP的cDNA， At28RNP 。叶绿体导入实验和RNA UV交联已经证实了该蛋白的推导序列表明其定位和RNA结合能力。 At28RNP 的表达分析表明，mRNA和编码的质体定位蛋白在活性组织中积累。 At28RNP蛋白与菠菜叶绿体蛋白So28RNP具有高度的序列同一性（60.6％），该蛋白与质体RNA的3 '末端结合，该质体RNA的非翻译区包含转录稳定的反向重复序列（IR） 。 体外 RNA加工试验表明，So28RNP对于将前体RNA正确加工成成熟形式是必需的。鉴定和分离同源蛋白n Arabidopsis 可以使用逆向遗传方法对质体局部RNP功能进行体内分析。转基因的拟南芥植物产生的At28RNP水平升高和降低，并且蛋白质水平在两种类型的转基因植物中均受到显着影响，但就质体影响而言，转基因植物与对照之间没有发现显着差异RNA成熟或积累。然而，转基因品系中改变的At28RNP蛋白表达的稳定遗传可能被证明可用于进一步研究叶绿体基因表达中的RNP。

著录项

  • 作者

    Phayre, Dina Jean.;

  • 作者单位

    University of California, Berkeley.;

  • 授予单位 University of California, Berkeley.;
  • 学科 Biology Microbiology.
  • 学位 Ph.D.
  • 年度 2000
  • 页码 p.3410
  • 总页数 217
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 微生物学;
  • 关键词

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