SGER: Synthetic Genes for the Design of Hydroxproline-Rich Glycomodules

SGER：用于设计富含羟脯氨酸的糖模块的合成基因

• 批准号: 9805960
• 负责人: Marcia Kieliszewski
• 金额: $ 5万
• 依托单位: Ohio University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-15 至 1999-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9805960&HistoricalAwards=false
• 关键词: SGER; Synthetic; Genes; Design; Hydroxproline

9805960 Kieliszewski Hydroxyproline-rich glycoproteins (HRGPs) are ubiquitous architectural components of growing plant cell walls, often accounting for 10-20% of their dry weight; polysaccharides account for the bulk. HRGPs are implicated in all aspects of plant growth and development including responses to stress. The HRGP superfamily contains three major groups representing a continuum of glycosylation and periodicity. They range from the repetitive proline-rich proteins that are lightly glycosylated and highly periodic, through the crosslinked extensins that are highly arabinosylated and periodic, to the arabinogalactan-proteins that are the most highly glycosylated and least periodic. The repetitive units are small, often only 4-6 residue glycosylated motifs viewed hypothetically as functional modules. The function of these unique plant glycoproteins remains largely unexplored. We propose to define their roles through functional analysis of single glycopeptide modules when expresssed as repetitive glycopolypeptides in tomato cultures. We shall insert synthetic genes encoding HRGP glycopeptide modules into two plant transformation vectors both derived from pBI121; one with a reporter gene, the other, for test for module function, lacking the reporter. We shall use these plasmids to transform cell cultures of tomato, which contain the plant prolylhydroxylases and glycosyltransferases required for module posttranslational modifications. Expression and isolation of these modules will enable us to test general and specific hypotheses for glycosylation and molecular function: 1. Glycosylation codes predicted by the Hyp-contiguity hypothesis specify precise saccharide addition to specific HRGP Hyp residues, namely: tetraarabinose to the most highly contiguous residues, arabinosylation decreasing with decreasing Hyp-contiguity; and polygalactosylation of repetitive noncontiguous Hyp residues that typify AGPs. 2. Carbohydrate-mediated alignment of extensin monomers is necessary for their orderly assembly and crosslinking roles in cell wall self-assembly.

9805960 Kieliszewski羟基羟基糖蛋白（HRGPS）是生长植物细胞壁的无处不在的建筑成分，通常占其干重的10-20％；多糖占批量。 HRGP与植物生长和发育的各个方面有关，包括对压力的反应。 HRGP超家族包含三个主要组，代表糖基化和周期性的连续体。 它们的范围从易于糖基化且高度周期性的富含脯氨酸的蛋白质到高度阿拉伯蛋白酶基化且周期性的交联延伸蛋白，到阿拉伯糖基乳糖蛋白，这些蛋白是最高度高度糖基化且最少周期性的蛋白。 重复单元很小，通常只有4-6个残基糖基化基序被认为是功能模块。 这些独特的植物糖蛋白的功能在很大程度上尚未探索。 我们建议通过对番茄培养物中的重复糖糖肽的单个糖肽模块的功能分析来定义它们的作用。 我们将插入编码HRGP糖肽模块的合成基因中的两个植物转化载体均衍生出PBI121。一个带有报告基因的基因，另一个用于模块函数的测试，缺乏记者。 我们将使用这些质粒转化番茄的细胞培养物，番茄中含有植物性丙基羟基酶和模块翻译后修饰所需的糖基转移酶。 这些模块的表达和隔离将使我们能够测试糖基化和分子功能的一般和特定假设：1。由催眠率假设假设预测的糖基化代码精确地添加了特定的HRGP炒作，即在特定的HRGP炒作中，即与高度偏置的降低了非依赖性的降低酶，阿非元素降低了非易位的降低。以及代表AGP的重复性非连续性催眠残基的多边形化。 2。碳水化合物介导的延伸素单体的比对，对于它们在细胞壁自组装中的有序组装和交联作用是必需的。