首页> 外文期刊>DNA Sequence: The Journal of DNA Sequencing and Mapping >Alternative splicing and expression analysis of OsFCA (FCA in Oryza sativa L.), a gene homologous to FCA in Arabidopsis
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Alternative splicing and expression analysis of OsFCA (FCA in Oryza sativa L.), a gene homologous to FCA in Arabidopsis

机译:拟南芥中与FCA同源的基因OsFCA(水稻中的FCA)的可变剪接和表达分析

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A subtracted cDNAs library was constructed using rice (Oryza sativa L.) calli cDNA as driver and differentiating calli cDNA as tester. A novel gene homologous with FCA in Arabidopsis was cloned from rice by screening the SSH (suppression subtractive hybridization) library followed by RACE. Four alternative transcripts of OsFCA were cloned from the leaves of rice, and designated as OsFCA-1, OsFCA-2, OsFCA-3 and OsFCA-4 respectively. OsFCA-1 was homologous to FCA- gamma of Arabidopsis and contained several conserved domains (two RNA Recognition Motifs and one WW-domain). OsFCA-2 was 102 bp shorter than OsFCA-1 which caused the WW-domain deletion. The proteins encoded by OsFCA-3 and OsFCA-4 were 101 amino acids shorter than OsFCA-1 at the N-terminal which is a glycine-rich region. The fluorescence quantitative PCR analysis showed that the mRNA of OsFCA-1 is the most abundant in the four splicing variants of rice FCA, and its expression level is much higher in differentiating calli than in calli. The expression of OsFCA-1 is steady in the leaves of three different stage, but up-regulated in young spikelet of primary branch-differentiating stage and down-regulated in young spikelet of pistil and stamen-differentiating stage.
机译:以水稻(Oryza sativa L.)愈伤组织cDNA为驱动基因,以分化愈伤组织cDNA为检测基因,构建了一个减法cDNA文库。通过筛选SSH(抑制消减杂交)文库,然后筛选RACE,从水稻中克隆了一个与拟南芥中FCA同源的新基因。从水稻叶片中克隆了四个OsFCA转录本,分别命名为OsFCA-1,OsFCA-2,OsFCA-3和OsFCA-4。 OsFCA-1与拟南芥的FCA-γ同源,并包含几个保守域(两个RNA识别基元和一个WW域)。 OsFCA-2比导致WW域删除的OsFCA-1短102 bp。 OsFCA-3和OsFCA-4编码的蛋白质在N端(富含甘氨酸的区域)比OsFCA-1短101个氨基酸。荧光定量PCR分析表明,OsFCA-1的mRNA在水稻FCA的四个剪接变体中含量最高,并且在分化愈伤组织中的表达水平远高于愈伤组织。 OsFCA-1的表达在三个不同阶段的叶片中稳定,但在初生枝分化期的幼小穗上调而在雌蕊和雄蕊分化期的幼小穗中下调。

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