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Cloning and Expression Analysis of Wheat Cytokinin Oxidase/Dehydrogenase Gene TaCKX3

机译:小麦细胞分裂素氧化酶/脱氢酶基因TaCKX3的克隆及表达分析

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摘要

Cytokinin oxidase/dehydrogenase plays an important role in regulating plant growth and development. A novel gene of TaCKX3 was cloned from wheat by specific primers designed according to the Triticeae Full-Length CDS Database and was proved to be located on chromosome 7B by analyzing the nulli-tetrasomic lines of "Chinese Spring." The genomic coding sequence was interrupted by three introns, which were 103, 421, and 458 bp, successively. The complementary DNA sequence of TaCKX3 is 60% identical to that of TaCKX1, and the homology of their deduced proteins is even low. The putative TaCKX3 protein shows highly homology with ZmCKX10 but not with other known cytokinin oxidase/dehydrogenase. However, it contains conserved motifs as other cytokinin oxidase/dehydrogenase, such as flavin adenosine dinucleotide binding domain and cytokinin binding domain. Consistent with ZmCKX10 and AtCKX7, nor the putative TaCKX3 has signal peptide at N terminus, which means that TaCKX3 functions in cytoplasm. Quantitive polymerase chain reaction analysis indicated that the expression of TaCKX3 gene is significantly up-regulated in germinating embryos treated by 6-BA and slightly up-regulated by NaCl or PEG-6000.
机译:细胞分裂素氧化酶/脱氢酶在调节植物生长发育中起重要作用。通过根据小麦的全长CDS数据库设计的特异性引物,从小麦中克隆了TaCKX3的新基因,并通过分析“中国春”的无效四体性系证明其位于7B染色体上。基因组编码序列被三个内含子(分别为103、421和458 bp)中断。 TaCKX3的互补DNA序列与TaCKX1的DNA序列具有60%的同一性,其推导蛋白质的同源性甚至很低。推定的TaCKX3蛋白与ZmCKX10具有高度同源性,但与其他已知的细胞分裂素氧化酶/脱氢酶没有高度同源性。但是,它含有保守的基序,如其他细胞分裂素氧化酶/脱氢酶,例如黄素腺苷二核苷酸结合结构域和细胞分裂素结合结构域。与ZmCKX10和AtCKX7一致,推定的TaCKX3也不在N末端具有信号肽,这意味着TaCKX3在细胞质中起作用。定量聚合酶链反应分析表明,TaCKX3基因的表达在6-BA处理的发芽胚中显着上调,而在NaCl或PEG-6000中略微上调。

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