园艺学报 ›› 2017, Vol. 44 ›› Issue (6): 1181-1188.doi: 10.16420/j.issn.0513-353x.2016-0708

• 研究报告 • 上一篇    下一篇


肖旭峰1,张 祎1,吴智明2,杨寅桂1,*   

  1. (1江西农业大学农学院,南昌 330045;2仲恺农业工程学院,广州 510225)
  • 出版日期:2017-06-25 发布日期:2017-06-25

Cloning,Sequence Analysis and Expression Profile of Protein Arginine N-methyltransferase Gene BrcuPRMT5 in Flowering Chinese Cabbage

XIAO Xufeng1,ZHANG Yi1,WU Zhiming2,and YANG Yingui1,*   

  1. (1College of Agronomy,Jiangxi Agricultural University,Nanchang 330045,China;2College of Horticulture and Landscape Architecture,Zhongkai University of Agriculture and Engineering,Guangzhou 510225,China)
  • Online:2017-06-25 Published:2017-06-25


 以‘油青49’和‘油青甜菜薹80’菜薹茎尖为材料,采用RT-PCR和RACE技术克隆获得组蛋白甲基转移酶基因BrcuPRMT5的全长cDNA和gDNA序列。BrcuPRMT5 cDNA序列全长为2 117 bp,其中完整开放阅读框为1 929 bp,编码642个氨基酸,相对分子量为71.55 kD,理论等电点(pI)为5.87;多序列比对结果表明,BrcuPRMT5编码的氨基酸序列含有高等植物PRMT5基因1个高度保守的结构域;系统发育分析结果显示与大白菜、油菜及甘蓝的亲缘关系最近;亚细胞定位软件分析得知,BrcuPRMT5蛋白无跨膜区域,可能定位于线粒体中;对应的gDNA全长为4 151 bp,含有23个外显子和22个内含子,最长的外显子长度为140 bp,内含子的长度范围为50 ~ 150 bp。利用半定量RT-PCR和实时荧光定量PCR技术分析基因的表达,BrcuPRMT5在菜薹不同组织中均有表达,其中在花中表达量最高,叶次之,根中最低;BrcuPRMT5从苗期至完全抽薹开花期的表达量呈现上升趋势。BrcuPRMT5在菜薹花发育过程中可能起一定的调控作用。

关键词: 菜薹, 蛋白甲基转移酶, 克隆, 序列分析, 时空表达

Abstract: The full length cDNA and gDNA of BrcuPRMT5 were obtained by using RT-PCR and RACE amplification from flowering Chinese cabbage(Brassica rapa syn. campestris ssp. chinensis var. utilis)‘Youqing 49’and‘Youqing Tiancaitai 80’stem apex. BrcuPRMT5 was 2 117 bp in full length and encoded a predicted protein of 642 amino acids(ORF length 1 929 bp)with a molecular mass of 71.55 kD and an isoelectric point of 5.87. Amino acid sequence analysis showed that it contains one conserved domains of PRMT family of protein arginine N-methyltransferase in higher plants. Phylogenetic tree analysis showed that the flowering Chinese cabbage had the closest evolutionary relationship with Chinese cabbage,rape and broccoli. Subcellular localization analysis indicated that it might be targeted to the mitochondrion without transmembrane region. The corresponding gDNA was 4 151 bp in length which harbored 23 exons and 22 introns. The longest extron was 140 bp in length and the length range of introns was 50–150 bp. BrcuPRMT5 was expressed in the root,stem,leaf and flower by the tissue-specificity expression. The expressive content in different tissues was different:the highest was in flower,the next was in stem,and the lowest level was in root. The semi-quantitative RT-PCR and qPCR analysis revealed that no detectable levels were expressed during the first stage of sampling,then transcripts were detected during the two true-leaf,the four true-leaf,the five true-leaf and the flowering stages. This suggests BrcuPRMT5 may play an important role in flowering Chinese cabbage flower development.

Key words: flowering Chinese cabbage, protein arginine N-methyltransferase, clone, sequence analysis, expression