菜心组蛋白乙酰转移酶基因BrcuHAC1克隆与表达分析

发布时间：2018-08-04 13:38

【摘要】：为揭示组蛋白乙酰化修饰在十字花科作物抽薹发育表观遗传调控中的功能,以油青49和油青甜菜心80天的菜心为材料,通过PCR和RACE方法克隆了菜心BrcuHAC1基因c DNA和g DNA全长序列,并进行生物信息学分析,同时采用实时荧光定量PCR方法检测目的基因的时空表达特性。结果表明,克隆获得了BrcuHAC1基因全长6 061 bp,包括CDS区5 067 bp,编码1 688个氨基酸。对应的g DNA全长为7 376 bp,含有15个外显子和14个内含子,最长的外显子长度为1 599 bp,内含子的长度范围为50~210 bp。启动子序列中含有多个基本转录元件和顺式作用元件,如光反应元件、脱落酸响应元件、参与厌氧诱导的增强子元件等。多序列比对结果表明,BrcuHAC1的氨基酸序列中含有高等植物HAC1基因的6个保守结构域。系统发育分析结果显示,菜心与油菜、甘蓝、白菜处于同一分支,其亲缘关系最近。时空特异性分析结果表明,BrcuHAC1在菜心的根、茎、花、叶中均有表达,其中在花中表达量最高,叶次之,根最低;BrcuHAC1从苗期至完全抽薹开花期的表达量呈现上升趋势,说明BrcuHAC1在菜薹花发育过程中可能起到一定的调控作用。本研究结果为揭示组蛋白乙酰转移酶调控菜心抽薹分子机制提供了一定的理论依据。
[Abstract]:In order to reveal the function of histone acetylation modification in epigenetic regulation of bolting and development of cruciferous crops, the full-length sequences of c DNA and g DNA of BrcuHAC1 gene were cloned by PCR and RACE. Bioinformatics analysis and real-time fluorescence quantitative PCR were used to detect the temporal and spatial expression characteristics of the target gene. The results showed that the full length of BrcuHAC1 gene was 6 061 BP, including CDS region 5 067 BP, encoding 1 688 amino acids. The total length of g DNA is 7 376 BP, which contains 15 exons and 14 introns. The longest exon length is 1 599 BP, and the length range of intron is 50 ~ 210bp. The promoter sequence contains a number of basic transcription elements and cis-acting elements, such as photoreactive elements, abscisic acid response elements, anaerobic-induced enhancers, and so on. The results of multiple sequence alignment showed that the amino acid sequence of BrcuHAC1 contained six conserved domains of HAC1 gene of higher plants. Phylogenetic analysis showed that Brassica rapeseed, Brassica napus and Brassica campestris were in the same branch and had the closest phylogenetic relationship. The results of spatio-temporal specificity analysis showed that BrcuHAC1 was expressed in root, stem, flower and leaf of Chinese cabbage, among which the highest expression was in flower, followed by leaf, and the lowest BrcuHAC1 in root showed an increasing trend from seedling stage to full bolting stage. The results suggest that BrcuHAC1 may play a regulating role in the development of cauliflower. The results provide a theoretical basis for revealing the molecular mechanism of histone acetyltransferase regulating bolting of Chinese cabbage.
【作者单位】： 江西农业大学农学院;
【基金】：国家自然科学基金(31360484) 教育部高等学校博士学科点专项科研基金(20133603120006) 江西省自然科学基金项目(2013BAB204009)
【分类号】：Q943.2;S634.5
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本文编号：2164053