Isolation and Expression Analysis of Fructokinase Genes from Citrus

文献类型： 外文期刊

第一作者： Qin, QP

作者： Qin, QP;Zhang, SL;Chen, JW;Xie, M;Jin, YF;Chen, KS;Asghar, S

作者机构：

关键词： citrus;fructokinase;gene cloning;expression analysis;fructose accumulation

期刊名称：ACTA BOTANICA SINICA （ 影响因子：0.599； ）

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年卷期：

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收录情况： SCI

摘要： Fructokinase (FRK) is of primary importance in phosphorylation of fructose in plants. Two genomic DNA fragments encoding putative fructokinase gene were isolated from Citrus unshiu Marc, using PCR, which were named Cufrk1 and Cufrk2. A cDNA sequence same to the exons' sequences of Cufrk1 was amplified by RT-PCR and the full-length cDNA sequences of this gene was isolated using rapid amplification of cDNA ends (RACE) from mature fruit, named CuFRK1 (GenBank number: AY561840). Sequencing analysis showed that the amino acid sequences were 68% identical between Cufrk1 and Cufrk2. The full-length cDNA of CuFRK1 was 1 459 bp in length and contained a complete ORF from 168 to 1 220 bp, encoding 350 amino acids with a calculated molecular weight of 37.5 kD and a predicted pI of 5.03. The deduced amino acids of CuFRK1 possessed two sugar-binding domains, three ATP-binding domains and were 62% to 78% identical to the previously characterized fructokinase genes of other plants. Northern analysis showed that transcripts of CuFRK1 (Cufrk1) and Cufrk2 were detected at a high level in leaves, fruit at early developmental stages, but undetectable in peels and stems. Their expression patterns were distinct in petals and mature fruit. Enzymatic analysis showed that the activity of citrus fructokinase was decreasing with the fruit development, coincided with the accumulation of fructose in fruit. There was a significantly negative correlation between the decreasing fructokinase activity and the increasing fructose content during fruit development.

分类号： Q94

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