天山雪莲sikFBA1基因克隆定位及表达分析
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国家自然科学基金(31360053)


Cloning, Subcellular Localization and Expression Analysis of sikFBA1 from Saussurea involucrata Kar. et Kir
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    摘要:

    果糖1,6二磷酸醛缩酶FBA家族(fructosel,6bisphosphate aldolase)在植物响应逆境调控中具有重要的意义。该研究基于天山雪莲低温转录组研究,采用RTPCR方法克隆了1个sikFBA1基因, ORF长度为1 077 bp,共编码358个氨基酸,含有1个保守的Glycolytic结构域,具有典型的FBA家族特征。系统进化分析发现,sikFBA1蛋白分类上属于Ⅰ型FBA,与来自四叶参(Codonopsis lanceolata)的同源蛋白亲缘关系最近。亚细胞定位结果表明,sikFBA1蛋白定位于细胞质,属于胞质型同工酶,与预测一致。实时定量PCR结果显示,天山雪莲经过不同低温(4 ℃/-2 ℃)处理,sikFBA1基因的表达水平整体下调,但该基因在冷胁迫与冰冻胁迫、冷驯化与非冷驯化下呈现出不同的表达模式。研究表明,sikFBA1基因参与了天山雪莲低温胁迫的响应。

    Abstract:

    The fructose1, 6bisphosphate aldolase (FBA) family plays an important role in plants stressresponse. A sikFBA1 gene was cloned based on the sequencing results of the Saussurea involucrata Kar. et Kir transcriptome with RTPCR method. The open reading frame (ORF) of the sikFBA1 gene is 1 077 bp which encoded a deduced protein including 358 amino acids residues. The protein sequence of sikFBA1 possesses the conserved Glycolytic domain, the typical family characteristic of FBA. Phylogenetic analysis exhibitied that sikFBA1 of S. involucrata Kar. et Kir showed the closest genetic relationship with that of Codonopsis lanceolata and they are all belonged to the FBA class I .When fused to green fluorescent protein (GFP) in the subcellular localization assay, we found that sikFBA1 protein was present in the cytoplasm, which was consistent with the predicted results. sikFBA1 is cytoplasmic isoenzyme. Quantitative realtime PCR (qRTPCR) was performed to determine the expression pattern in 4 ℃ and -2 ℃ during the low temperature treatment. The results showed that the expression level of sikFBA1 gene was generally downregulated after treatment. Interestingly, the sikFBA1 gene showed different expression patterns between the different stress conditions of cold and freeze, cold acclimation and unacclimation respectively. These results indicated that sikFBA1 had the potential function in S. involucrata Kar. et Kir response to low temperature stress.

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穆建强,梁文洁,张芳松,等.天山雪莲sikFBA1基因克隆定位及表达分析[J].西北植物学报,2016,36(11):2137-2145

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  • 在线发布日期: 2016-12-12
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