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菜心 BclUBE2 基因的克隆与表达分析
引用本文:曾小玲,赵瑞丽,钟开勤,朱朝辉,陈敏氡.菜心 BclUBE2 基因的克隆与表达分析[J].热带作物学报,2018,39(9):1722-1777.
作者姓名:曾小玲  赵瑞丽  钟开勤  朱朝辉  陈敏氡
作者单位:1福州市蔬菜科学研究所,福建福州 350111; 2. 福建省农业科学研究院作物研究所
摘    要:采用同源克隆法从菜心中获得 cDNA 全长 459 bp 的 E2 基因,命名为 BclUBE2。该基因编码 153 个氨基酸,分子量为 17.24 ku,理论等电点为 5.37,为亲水性非分泌蛋白。结构分析显示,该蛋白含有一个泛素结合酶E2 活性位点、一个 WD 重复序列和一个高度保守的半胱氨酸。进化分析发现,菜心 BclUBE2 蛋白与芸薹属芜菁和油菜的亲缘关系较近。qRT-PCR 分析表明,菜心 BclUBE2 基因在根、茎、叶、叶柄中均有表达,且表达丰度和变化趋势不同。在低温胁迫下,根中的表达量呈现先降低后升高的变化趋势,且在处理 1 h 时表达量最低;在茎、叶、叶柄中均呈现先升高后降低的表达趋势,茎和叶柄处理 6 h 时的表达量最高,叶片处理 1 h 时的表达量最高。说明菜心 BclUBE2 基因在响应低温胁迫中发挥作用。

关 键 词:菜心  BclUBE2基因  qRT-PCR  

Cloning and Expression Analysis of Ubiquitin-Conjugating Enzyme BclUBE2 Gene in Brassica campestris L. ssp. chinensis var. utilis
ZENG Xiaoling,ZHAO Ruili,ZHONG Kaiqin,ZHU Chaohui,CHEN Mindong.Cloning and Expression Analysis of Ubiquitin-Conjugating Enzyme BclUBE2 Gene in Brassica campestris L. ssp. chinensis var. utilis[J].Chinese Journal of Tropical Crops,2018,39(9):1722-1777.
Authors:ZENG Xiaoling  ZHAO Ruili  ZHONG Kaiqin  ZHU Chaohui  CHEN Mindong
Institution:1Fuzhou Institute of Vegetable Science, Fuzhou, Fujian 350111, China;  2. Crops Research Institute, Fujian Academy of Agricultural Sciences,
Abstract:The ubiquitin-conjugating enzyme E2 gene was cloned from Brassica campestris L. ssp. chinensis var. utilis Tsen et Lee and designated as BclUBE2. The full cDNA length of BclUBE2 was 459 bp, encoding 153 amino acid residues with an estimated molecular mass of 17.24 ku and a calculated pI of 5.37. This protein was hydrophilic non-secretory. Structural analysis revealed that the sequence contained an E2 ubiqitin-conjugating enzyme active site, a WD-repeated sequence and a highly conserved cysteine. Phylogenetic analysis showed that BclUBE2 protein had the closest relationship with Brassica napus and Brassica rapa. qRT-PCR results showed that the existence of BclUBE2 was detectable in the root, stem, leaf and petiole with different expressive abundance and change trend. Under low temperature stress, the expression level in the root showed the trend of decreasing first and then increasing, and reached the lowest level at 1 h. However, the expression level in the stem, leaf and petiole increased first and then decreased with the highest level at 6 h in stem and petiole and at 1 h in leaf. These results indicated that BclUBE2 gene was involved in plant response to low temperature stress.
Keywords:Brassica campestris L  ssp  chinensis var  utilis Tsen et Lee  BclUBE2 gene  qRT-PCR  
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