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| 大麦(Hordeum vulgare)昼夜节律钟基因CCA1的克隆及表达分析 | |
| 引用本文： | 邢国芳,宋萌,姚涵,韩渊怀.大麦(Hordeum vulgare)昼夜节律钟基因CCA1的克隆及表达分析[J].山西农业大学学报(自然科学版),2012,32(3):207-212. |
| 作者姓名： | 邢国芳 宋萌 姚涵 韩渊怀 |
| 作者单位： | 1. 山西农业大学农学院,山西太谷030801;山西农业大学生物工程研究所,山西太谷030801 2. 山西运城农业职业技术学院,山西运城,044000 3. 山西农业大学农学院,山西太谷,030801 |
| 基金项目： | 山西农业大学科技创新基金，山西农业大学博士科研启动基金 |
| 摘 要： | 昼夜节律钟基因CCA1在调解水稻和拟南芥的光周期反应中起着重要作用。利用BLAST手段以玉米中的CCA1基因序列作为靶序列,调取Genbank数据库信息,并结合RT-PCR方法获得了大麦的cDNA同源序列。BLASTx分析发现其与水稻和玉米的序列相似性分别达到72%和69%。通过ORF Finder软件分析发现,该序列包含一个2157bp的开放阅读框,编码一个由718个氨基酸残基组成的蛋白序列,其分子量为77 769.4Da,等电点为6.55。采用实时荧光定量PCR分析发现,随光照时间的变化,该基因在大麦叶片中的表达量呈现出白天不断降低而夜晚逐渐升高的昼夜变化趋势,且在华大麦1号和华大麦2号中的表达曲线存在一定的差异。本研究为进一步研究大麦CCA1基因在调控大麦光周期响应途径中的功能,阐明大麦光周期敏感机制提供了科学依据。 |
| 关 键 词： | 光周期 基因克隆 基因表达 BaCCA1 |
Cloning and Expression Analysis of Circadian Clock Gene CCA1 in Barley (Hordeum vulgare) |
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| Xing Guofang , Song Meng , Yao Han , Han Yuanhuai.Cloning and Expression Analysis of Circadian Clock Gene CCA1 in Barley (Hordeum vulgare)[J].Journal of Shanxi Agricultural University,2012,32(3):207-212. | |
| Authors: | Xing Guofang Song Meng Yao Han Han Yuanhuai |
| Institution: | 1,2(1.College of Agriculture,Shanxi Agricultural University,Taigu Shanxi 030801,China;2.Agricultural Bioengineering Center,Shanxi Agricultural University,Taigu Shanxi 030801,3.Institute of Yuncheng Agricultural Professional Technology,Yunchen Shanxi 044000) |
| Abstract: | CCA1 gene plays an important role in circadian clock sensitivity in rice(Oryza sativa L.) and Arabidopsis thaliana.In this study,CCA1 gene in barley was cloned by RT-PCR using homological primers based on the highly conserved region of the multiple alignments of the rice and Arabidopsis.The similarities of this sequence were up to 72% and 69%,respectively,to corresponding mRNA sequences of rice and maize in BLASTx of GenBank database.Using ORF Finder software,a 2157 bp open reading frame was found to code 718 amino acids.Using Compute pI/Mw tool,the amino acid sequence was analyzed,and it revealed that the molecular weight of this protein was about 77 769.4 Da,and isoelectric point was about 6.55.We established fluorescence quantitative RT-PCR system with barley inbred lines HUADAMAI 1 and HUADAMAI 2,and studied the expression of CCA1 in leaf under 16h/8h(light/dark) conditions.Expression analysis showed that the gene expression peaked at dawn(ZT0) then gradually declined from ZT0 to ZT15,bottomed at ZT15,then increased and returned to the initial level at ZT24.This study will provide information of barley CCA1 gene for further studying the function in regulating photoperiod sensitivity in barley,and provide scientific basis for clarifying the mechanism of the circadian synchronization in barley. |
| Keywords: | Photoperiod Gene cloning Gene Expression BaCCA1 |
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