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Isolation and characterization of the gene for the biosynthesis of the rice storage protein using the mutants

使用突变体分离和表征水稻贮藏蛋白生物合成基因

基本信息

批准号：
10660009
负责人：
KUMAMARU Toshihiro
金额：
$ 2.3万
依托单位：
KYUSHU UNIVERSTIY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

项目摘要

・ prolamin regulatory gene mutantThe expression pattern of mRNA from the immature seeds of the prolamin mutant, which showed the decreased level of several prolamin molecules, was analyzed by the differential display methods. The expression pattern of some bands in mutants differed from that of the original variety.・ The mutant for the transportation and accumulation of glutelinProglutelin precursor high accumulation mutant (57H), esp2 mutant, lacked protein disulfide isomerase (PDI), one of the molecular chaperon, at both the RNA and protein levels. These results suggest that PDI plays and essential role in the segregation of proglutelin and prolamin polypeptides within the ER lumen. The Esp2 gene differed from the gene coding the PDI and the other esp2 mutant line also showed depressed PDI mRNA level. This suggests that the Esp2 gene codes the gene activator protein of PDI gene. The deduced amino acid sequence of the partially isolated PDI shows that rice sequence identity to maize, barley, wheat and castor bean clones is 84.8%, 84.2%, 83.9% and 62.5%, respectively.・ glutelin structural gene mutantThe genetic analyses show that three types of glutelin α-2 subunit lacking mutants (α-2L mutant) were the mutants of the same locus for the structural gene of glutelin. The gene of α-2L mutant was named as Glu4,. Which was corresponded to cDNA clone GluA1.

·醇溶谷蛋白调节基因突变体通过差异显示方法分析了来自醇溶谷蛋白突变体的未成熟种子的mRNA的表达模式，所述突变体显示了几种醇溶谷蛋白分子的水平降低。突变体中某些条带的表达模式与原始品种不同。·谷蛋白前体谷蛋白高积累突变体（57 H），特别是突变体2，在RNA和蛋白质水平上均缺乏分子伴侣蛋白质二硫键异构酶（PDI）。这些结果表明，PDI在内质网腔内谷蛋白原和醇溶蛋白多肽的分离中起着重要作用。Esp 2基因与编码PDI的基因不同，并且另一个esp 2突变体系也显示出抑制的PDI mRNA水平。这表明Esp 2基因编码PDI基因的基因激活蛋白。推导的氨基酸序列表明，水稻与玉米、大麦、小麦和蓖麻克隆的同源性分别为84.8%、84.2%、83.9%和62.5%。谷蛋白结构基因突变体的遗传分析表明，谷蛋白α-2亚基缺失突变体（α-2L突变体）是谷蛋白结构基因的同一位点突变体。将α-2L突变体的基因命名为Glu 4。该基因与cDNA克隆GluA 1相对应。