Ripening-related occurrence of phosphoenolpyruvate carboxykinase in tomato fruit
Ripening-related occurrence of phosphoenolpyruvate carboxykinase in tomato fruit
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DOI：
10.1023/a:1011842828723
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发表时间：
2001-01-01
影响因子：
5.1
通讯作者：
Gray, JE
中科院分区：
文献类型：
--
作者：
Bahrami, AR;Chen, ZH;Gray, JE
磷酸烯醇式丙酮酸羧激酶（PEPCK）存在于成熟的番茄果实中。从成熟番茄果实的cDNA文库的PCR筛选中鉴定了编码PEPCK的cDNA。番茄PEPCK cDNA的序列和基因组DNA的克隆部分显示，完整的cDNA序列包含编码长度为662个氨基酸残基的肽的开放阅读框，并预测分子量为73.5 kDa的多肽，其对应于通过蛋白质印迹检测的多肽。在番茄基因组中仅鉴定出一个PEPCK基因。通过活性测定、蛋白质印迹和mRNA分析，证明PEPCK存在于成熟番茄果实的果皮中。PEPCK的丰度和活性都在果实成熟过程中增加，从未成熟绿色果实中检测不到的量到成熟果实中的高量。PEPCK的mRNA，蛋白质和活性也检测到萌发的种子，在较低的量，在番茄的根和茎。对PEPCK在番茄果实成熟过程中的作用进行了讨论。
Phosphoenolpyruvate carboxykinase (PEPCK) is present in ripening tomato fruits. A cDNA encoding PEPCK was identified from a PCR-based screen of a cDNA library from ripe tomato fruit. The sequence of the tomato PEPCK cDNA and a cloned portion of the genomic DNA shows that the complete cDNA sequence contains an open reading frame encoding a peptide of 662 amino acid residues in length and predicts a polypeptide with a molecular mass of 73.5 kDa, which corresponds to that detected by western blotting. Only one PEPCK gene was identified in the tomato genome. PEPCK is shown to be present in the pericarp of ripening tomato fruits by activity measurements, western blotting and mRNA analysis. PEPCK abundance and activity both increased during fruit ripening, from an undetectable amount in immature green fruit to a high amount in ripening fruit. PEPCK mRNA, protein and activity were also detected in germinating seeds and, in lower amounts, in roots and stems of tomato. The possible role of PEPCK in the pericarp of tomato fruit during ripening is discussed.