中国水仙NtSOC1基因克隆及亚细胞定位
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国家自然科学基金(31601783);


Cloning and Subcellular Localization of NtSOC1 Gene from Narcissus tazetta var. chinensis
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    摘要:

    以中国水仙‘金盏银台’为实验材料,采用RACE和RTPCR技术获得1个与开花相关的转录因子(SOC1)的同源基因NtSOC1。NtSOC1的cDNA全长1 603 bp,含有1个687 bp开放阅读框,编码228个氨基酸。生物信息学分析表明,NtSOC1与单子叶植物的SOC1同源基因的氨基酸序列较为相似,且在C末端同样含有一个保守性很高的SOC1 motif序列,说明NtSOC1是属于SOC1/TM3亚家族基因。荧光定量PCR分析显示,NtSOC1在花芽分化阶段的表达量随着花芽的分化而升高,花芽分化结束时减少,表明NtSOC1基因可能参与中国水仙的花芽分化。成功构建了NtSOC1基因表达载体pCAMBIA1302NtSOC1,通过农杆菌转化洋葱表皮对编码蛋白进行亚细胞定位结果显示,NtSOC1基因编码蛋白定位于细胞核,符合转录因子的亚细胞定位特征。该实验结果为进一步研究NtSOC1基因的生物学功能奠定了基础。

    Abstract:

    A flowering regulated transcription factor SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) homologous gene, NtSOC1, was cloned from the flower bud of Narcissus tazetta var. chinensis using RTPCR and rapid amplification of cDNA ends. NtSOC1 is 1 603 bp in length with an open reading frame 687 bp which is capable of encoding 228 amino acids. Phylogenetic analysis indicated that the amino acid sequence of NtSOC1 is similar with the SOC1s in other species, especially the SOC1 in monocots. NtSOC1 contains a highly conservative SOC1 motif, which indicates that NtSOC1 belongs to the SOC1/TM3 subfamily gene. Fluorescence quantitative PCR analysis showed that the expression level of NtSOC1 in flower bud differentiation stage increased with the differentiation of flower bud and decreased at the end of flower bud differentiation, which indicated that NtSOC1 might be involved in flower bud differentiation of narcissus. The pCAMBIA1302NtSOC1 expression vector was successfully constructed. Recombinant plasmid was introduced into onion epidermal cells by Agrobacterium tumefaciens method. Subcellular localization result showed that the NtSOC1 was localized in the nucleus, which was consistent with the general character of subcellular localization of transcription factors. The results laid a foundation for further study of the function of NtSOC1 gene.

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武 丹,吴菁华,张志忠.中国水仙NtSOC1基因克隆及亚细胞定位[J].西北植物学报,2017,37(10):1889-1895

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  • 在线发布日期: 2017-12-01
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