水稻OsPDIL1-1 基因的功能初步分析
首发时间:2014-01-16
摘要:蛋白质二硫键异构酶(Protein Disulfide Isomerase,PDI)在衣藻及动物中研究较为详细,除人源典型结构PDI和人源胰腺PDI 外,Rb60、ERp57和ERp72在植物体中都有相似结构的蛋白。本研究通过图位克隆的方法在水稻中分离了一个水稻中控制淀粉合成的基因Protein Disulfide Isomerase Like 1-1 (OsPDIL1-1)。转基因互补结果进一步证实了图位克隆的结果。RT-PCR和western杂交结果显示PDIL1-1在水稻根、茎、叶、叶鞘、穗和不同发育时期的胚乳均有表达,在发育12天的胚乳中表达量最高,而突变体中则没有检测到PDIL1-1的表达。体外实验证实PDIL1-1蛋白具有还原酶活性及分子伴侣活性。荧光定量RT-PCR检测发现内质网胁迫相关基因的表达均被显著上调,同时PCD标志基因HSR203J在T3612中被诱导表达。因此PDIL1-1的缺失导致胚乳细胞发生严重的内质网胁迫,并极有可能启动PCD,最终影响淀粉的合成。
关键词: 水稻 粉质 内质网胁迫 蛋白质二硫键异构酶 功能分析
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Preliminary research on the function of OsPDIL1-1 in rice(Oryza sativa L.)
Abstract:Protein disulfide isomerase (PDI) study in more clear in Chlamydomonas and animals, Rb60, ERp57 and ERp72 in plants have a similar structure proteins except the typical structure of anthropogenic PDI and the anthropogenic pancreas PDI. This study separated a control rice starch synthesis genes Protein Disulfide Isomerase Like 1-1 (OsPDIL1-1) in rice by using the method of map-based cloning. Semi-quantitative RT-PCR and western blot showed PDIL1-1 in rice roots, stems, leaves, leaf sheath, panicle and different developmental stages of endosperm, and the expression amount of the highest in the developing endosperm of 12 days , while the mutants are not detected the PDIL1-1 expression. In vitro experiments have established that PDIL1-1 protein with reductase activity and molecular chaperone activity. Quantitative RT-PCR test found that the expression of related genes were endoplasmic reticulum stress rise significantly, at the same time PCD marker gene HSR203J in T3612 were induced to express. Therefore PDIL1-1 the lack of endosperm cells leads to serious endoplasmic reticulum stress, and more than likely start PCD, ultimately affect the synthesis of starch.
Keywords: Rice Floury ER stress Protein disulfide isomerase;Gene function analysis
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