菜薹蛋白精氨酸甲基转移酶基因BrcuPRMT5的克隆及表达分析

发布时间：2018-05-28 23:48

  本文选题：菜薹 + 蛋白甲基转移酶　； 参考：《园艺学报》2017年06期

【摘要】：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Brcu PRMT5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(p I)为5.87;多序列比对结果表明,Brcu PRMT5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,Brcu PRMT5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。
[Abstract]:The stem tips of 'Youqing 49' and 'you Qing sweet' cabbage bolting 80 'were used as materials. The full length cDNA and gDNA sequence. BrcuPRMT5 cDNA sequence of histone methyltransferase gene Brcu PRMT5 were cloned by RT-PCR and RACE techniques. The complete open reading frame was 1 929 BP, encoding 642 amino acids. The relative molecular weight was 71.55 KD and the theoretical isoelectric point I) was 5.87. The results of multiple sequence alignment showed that the amino acid sequence encoded by Brcu PRMT5 contained a highly conserved domain of PRMT5 gene in higher plants, and phylogenetic analysis showed that the amino acid sequence of Brcu PRMT5 was similar to that of Chinese cabbage. The relationship between Brcu and Brcu in Brassica napus was very close. The subcellular localization software showed that the Brcu PRMT5 protein was located in mitochondria and had no transmembrane region. The corresponding gDNA was 4 151 BP with 23 exons and 22 introns. The longest exon length is 140 BP, and the intron length range is 50 ~ 150 BP. The expression of BrcuPRMT5 was detected by semi-quantitative RT-PCR and real-time fluorescence quantitative PCR in different tissues of Chinese cabbage, in which the highest expression was found in flower, followed by leaf. The expression of BrcuPRMT5 increased from seedling stage to full bolting flowering stage. BrcuPRMT5 may play a regulatory role in the development of cauliflower.
【作者单位】： 江西农业大学农学院;仲恺农业工程学院;
【基金】：国家自然科学基金项目(31360484) 教育部高等学校博士学科点专项科研基金项目(20133603120006) 江西省自然科学基金项目(2013BAB204009) 江西农业大学博士启动基金项目(3181)
【分类号】：S634.5
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本文编号：1948667