蔷薇RrF3′H基因的克隆与分析

J. Jin, Zhongjian Li, Lan-yong Zhao
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引用次数: 1

摘要

蔷薇是蔷薇科蔷薇属的重要园林观赏植物。目前野生和栽培的野樱草多为紫色、粉红色,少量为白色,但缺乏黄色、橙色等。黄酮3′-羟化酶属于细胞色素P450的CYP75B亚家族,是花青素合成的必需酶。本实验利用RT-PCR技术从‘珲春’玫瑰花瓣中克隆rrf3’h基因,并进行生物信息学分析。rrf3’h基因开放阅读框全长1687 bp,编码510个氨基酸。RrF3'H编码的蛋白分子式为C2666H4149N699O734S24。所得蛋白分子量为58,506.95 Da。脂肪族指数为90.94。它属于不稳定的亲水蛋白。该蛋白由46.76%的α-螺旋、31.04%的随机螺旋、7.66%的β-角和14.54%的延伸链组成。该蛋白含有21个丝氨酸磷酸化位点,12个苏氨酸磷酸化位点和2个酪氨酸磷酸化位点。该蛋白含有两个o -糖基化位点,分别位于氨基酸序列的第98位和263位。该蛋白具有信号肽位点和跨膜结构。此外,通过比较rrf3’h的表达水平,我们发现rrf3’h与花的颜色深度呈正相关。
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Cloning and Analysis of RrF3’ H in Rosa rugosa
Rosa rugosa is an important garden ornamental plant which belongs to the genus Rosa of the family Rosaceae. The current wild and cultivated R. rugosa are mostly purple, pink, a small amount of white, but lack of yellow, orange and so on. Flavonoids 3’-hydroxylase belongs to CYP75B subfamily of cytochrome P450, and is an essential enzyme in anthocyanins synthesis. In this experiment, RrF3’H gene was cloned from the petal of Rosa rugosa ‘Hunchun’ using RT-PCR, and bioinformatics analysis was performed. The RrF3’H gene’s full length of opening reading frame was 1687 bp, encoding 510 amino acids. The formulas of proteins encoded by RrF3’H were C2666H4149N699O734S24. The derived protein had a molecular weight of 58,506.95 Da. The aliphatic index was 90.94. It belongs to unstable hydrophilic protein. The protein consists of 46.76% α-helix, 31.04% random coil, 7.66% β-corner and 14.54% extended strand. The protein contains 21 Ser phosphorylation sites, 12 Thr phosphorylation sites, and 2 Tyr phosphorylation sites. The protein contained two O-glycosylation sites, located at positions 98 and 263 of the amino acid sequence respectively. The protein has a signal peptide site and a transmembrane structure. In addition, by comparing the expression levels of RrF3’H, we found RrF3’H was positively correlated with the depth of flower color.
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