Cloning and Sequencing of a cDNA Clone Encoding the Cytosolic Triose-Phosphate Isomerase from Arabidopsis thaliana
Cloning and Sequencing of a cDNA Clone Encoding the Cytosolic Triose-Phosphate Isomerase from Arabidopsis thaliana
复制标题
编码拟南芥胞质磷酸丙糖异构酶的 cDNA 克隆的克隆和测序
DOI：
10.1104/pp.104.3.1103
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
M. Shih
中科院分区：
文献类型：
--
作者：
M. Shih
光合作用是将太阳能和CO2转化为碳水化合物的重要代谢途径之一。CO2的固定在叶绿体内通过主要由核基因编码的一组酶进行（Weeden和Gottlieb，1980; Cerff，1982; Shih等人，1986年）。除了光合固碳酶外，高等植物还具有一系列类似的酶，它们在胞质糖酵解中起作用。虽然这两种途径的酶功能非常相似，但它们参与碳代谢的相反方向。光合碳固定基因和糖酵解基因的表达受环境条件的不同影响（Yang et al.，1993年）。因此，这两组基因可以作为模式系统，用于研究环境条件的变化，如光，碳源和温度，如何影响植物的生长和发育在分子水平上。3-磷酸甘油酸转化为Fru-6-P的五个步骤对于两种途径是相同的，除了反应以相反的方向流动。涉及的叶绿体酶，磷酸果糖激酶、醛缩酶、TPI、甘油醛-3-P脱氢酶和磷酸甘油酸激酶，在结构和功能上与其胞质对应物非常相似（Weeden和Gottlieb，1980; Gottlieb，1982）。因此，编码这些酶的基因提出了非常有趣的模型，不仅用于研究编码叶绿体蛋白的核基因的进化，而且还用于研究参与两种主要碳代谢途径的基因在不同生理条件下是如何调节的。细胞溶质TPI的基因和cDNA克隆已从几种植物物种中获得，包括玉米（Marchionni和吉尔伯特，1986）和水稻（Xu和Hall，1993; Xu等人，1993年）。我在这里报告的克隆和表征（表一）的cDNA克隆编码胞质TPI从拟南芥。通过PCR扩增获得玉米TPI cDNA片段，并用于筛选拟南芥叶片cDNA文库。片段1序列分析和酶切图谱分析表明，分离的克隆属于相同的cDNA。测定最长克隆的完整核苷酸序列。假定的拟南芥胞质TPI克隆含有一个
Photosynthesis, which converts solar energy and COz into carbohydrates, is one of the most important metabolic pathways in plants. The fixation of C02 is camed out inside chloroplasts by a set of enzymes encoded mainly by nuclear genes (Weeden and Gottlieb, 1980; Cerff, 1982; Shih et al., 1986). In addition to photosynthetic carbon fixation enzymes, higher plants possess a set of similar enzymes that function in cytosolic glycolysis. Although the enzymic functions of the two pathways are quite similar, they are involved in the opposite directions of carbon metabolism. Expression of the photosynthetic carbon fixation genes and the glycolytic genes is affected differently by environmental conditions (Yang et al., 1993). Therefore, the two sets of genes can be used as model systems for studying how changing environmental conditions, such as light, carbon source, and temperature, affect plant growth and development at the molecular level. The five steps in the conversion of 3-phosphoglycerate into Fru-6-P are identical for the two pathways except that the reactions flow in opposite directions. The chloroplast enzymes involved, phosphofructokinase, aldolase, TPI, glyceraldehyde-3-P dehydrogenase, and phosphoglycerate kinase, are structurally and functionally very similar to their cytosolic counterparts (Weeden and Gottlieb, 1980; Gottlieb, 1982). Therefore, the genes encoding these enzymes present very interesting models for studying not only the evolution of nuclear genes encoding chloroplast proteins but also how genes involved in two primary carbon metabolic pathways are regulated under different physiological conditions. The genes and cDNA clones for cytosolic TPI have been obtained from severa1 plant species, including maize (Marchionni and Gilbert, 1986) and rice (Xu and Hall, 1993; Xu et al., 1993). I report here the cloning and characterization (Table I) of the cDNA clone encoding cytosolic TPI from Arabidopsis thaliana. A maize TPI cDNA fragment was obtained by PCR amplification and used to screen an Arabidopsis leaf cDNA library. Partia1 sequence analysis and restriction mapping indicated that isolated clones belong to the same cDNA. The complete nucleotide sequence for the longest clone was determined. The putative Arabidopsis cytosolic TPI clone contains an