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津田芜菁BrSIZ1基因克隆、定位及表达

         

摘要

SIZ1, a SUMO E3 ligase involved in post-translation of proteins in plant cells, plays a role in protein interaction, loca-tion, and response to environmental stresses. In order to elucidate the expression profile of SIZ1 in Tsuda, cDNA of SIZ1 gene was isolated from Tsuda. This gene was named BrSIZ1 (GenBank accession number KY441465). BrSIZ1 was 2754 bp in full length cDNA and 2571 bp in full length open reading frame (ORF), encoding a peptide with 856 amino acids. A BrSIZ1-GFP expression vector was constructed to analysis the subcellular localization. BrSIZ1-GFP was localized to nucleus, indicating that BrSIZ1 may play an important role in the nucleus. Quantitative-PCR analysis showed that the BrSIZ1 was expressed meetly in leaf and secondly in young seedling and red root epidermis, showing tissue specificity. The expression of the BrSIZ1 was induced by UV-A light in the root epidermis. The transcript level of BrSIZ1 was up-regulated when treated with temperature of 4℃or 37℃in young seedling.%SIZ1是植物细胞蛋白质翻译后修饰SUMO化的E3连接酶,参与植物蛋白相互作用、定位和抗逆反应.为研究BrSIZ1在津田芜菁中的表达特性,本研究克隆了津田芜菁SIZ1基因全长cDNA序列,命名为BrSIZ1(GenBank登录号为KY441465),该基因全长2754 bp,其ORF全长2571 bp,编码856个氨基酸残基的多肽.构建了BrSIZ1-GFP表达载体进行亚细胞定位研究,结果显示BrSIZ1-GFP定位于细胞核内,可能在细胞核中发挥其功能.利用荧光定量PCR检测表明,该基因表达量在叶片中最高,在幼苗和红色根皮中次之,表达具有组织特异性.而且BrSIZ1在芜菁根皮中的表达受长波紫外线(UV-A)诱导,在4°C、37°C胁迫的幼苗中,表达量增加.

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