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利用改进的DDRT-PCR分离并鉴定水稻细胞缺铁胁迫相关cDNA克隆

Cloning and Characterization of Novel Rice Cell cDNAs Under Iron-deficiency by Improved DDRT-PCR Method

  • 摘要: 利用改进的差异显示PCR(DDRT-PCR)技术从水稻细胞中分离与缺铁胁迫相关的新基因。在分离到的8个差异片段中,通过Northern杂交验证,其中5个为阳性。在这5个阳性片段中有3个是受缺铁胁迫抑制,有2个受缺铁胁迫诱导表达。对这些阳性片段的序列分析表明有3个(IDR2,IDR3,IDR8)为新基因。其中IDR2IDR8是2个未知基因,IDR3通过GenBank搜索可以找到一些与它同源的已知基因片段。

     

    Abstract: An improved differential display reverse transcript PCR(DDRT-PCR) approach was used to identify novel genes treated by iron-deficiency from rice cells.Total eight differential fragments(IDR1-8) were identified.Further northern blotting indicated that five of them were positive;three of those positive fragments were suppressed in iron-deficiency condition,while two of them were induced under iron-deficiency.Sequence analysis revealed that three of them (IDR2,IDR3,IDR8) represent novel genes.Two of them (IDR2,IDR8) were unknown genes,while the IDR3 was homogenous to some known genes by blasting approach.

     

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