摘要
本实验基于欧李转录组数据分析的基础上,利用RT-PCR方法从欧李品种‘农大4号’果实中克隆的到了ChCCD1基因,ChCCD1基因的CDS序列长度为1 644 bp,编码547个氨基酸的蛋白,分子量61.78 kD,等电点为6.05,不稳定指数为34.76。二级结构由α-螺旋,延伸链,β-折叠和随机卷须构成。蛋白三级结构建模分析发现,ChCCD1蛋白与玉米VP14具有相似的空间结构,具有7个由β-折叠组成的叶片,7个叶片中心含有4个组氨酸结合的Fe^(2+),构成了该酶的活性中心。通过同源氨基酸序列比对和进化树分析发现,ChCCD1编码的氨基酸序列与李和桃的CCD1基因相似性较高,达到99%。本研究为后续的基因的功能分析奠定了基础。
Based on the data analysis of Cerasus humilis transcriptome, this experiment used RT-PCR method to clone the ChCCD1 gene from the fruit of Cerasus humilis ‘Nongda 4 hao＇. The CDS sequence of ChCCD1 gene was 1 644 bp, encoding 547 amino acid proteins. The molecular weight was 61.78 kD, the isoelectric point was 6.05, and the unstable index was 34.76. The secondary structure consisted of α-helix, extended strand, β-turn and random coil. Protein tertiary structure modeling analysis found that ChCCD1 protein and Zea mays VP14 had similar spatial structure, which all contained 7 blades consisting of g-folding. 7 blade center contained Fe^2＋ united by 4 histidine, which constituted the active center of the enzyme. Through homologous amino acid sequence alignment and evolutionary tree analysis, we found that the amino acid sequence encoded by ChCCDI had high similarity with the CCD1 gene of Prunus rnume and Prunus persica, which reached 99%. This study laid the foundation for the further functional analysis of the gene.
作者
高利敏
王鹏飞
杜俊杰
穆霄鹏
王裕添
杜灵敏
张建成
Gao Limin ,Wang Pengfei, Du Junjie, Mu Xiaopeng, Wang Yutian, Du Lingmin ,Zhang Jiangcheng(College of Horticulture, Shanxi Agricultural University, Taigu, 03080)
出处
《分子植物育种》
CAS
CSCD
北大核心
2018年第5期1426-1432,共7页
Molecular Plant Breeding
基金
山西省自然科学基金项目(20120110032-4)
高等学校博士学科点专项科研基金项目(20101403120006)共同资助
关键词
欧李
类胡萝卜素裂解双加氧酶
生物信息分析
Cerasus humilis, Carotenoid cleavage dioxygenases, Bioinformatics analysis
