为更好地理解西红花类胡萝卜素合成代谢途径，从西红花中分离和克隆了1 个新的β-胡萝卜素羟化酶基因CsBCH1-z，并从柱头cDNA中克隆了该基因的全长编码序列为906 bp，编码301 个氨基酸残基，对应的基因组序列为1261 bp，5 个外显子由4 个内含子子间隔开。与已知的CsBCH1 基因的氨基酸序列比较，发现在N端由于6 个碱基的缺失，造成了2 个氨基酸的缺失和1 个氨基酸由异亮氨酸突变为亮氨酸，另外还有一处单碱基的变异造成苏氨酸突变为丙氨酸。通过预测其蛋白含有3 个明显的跨膜结构域，C端包含了2 组“HXXXXH”和“HXXHH”结构域，是典型的脂肪酸羟化酶。β-胡萝卜素羟化酶是西红花苷合成的重要前体物质，CsBCH1-z 柱头中的表达量>叶片>花瓣；雄蕊中表达很低；球茎中表达不可见。
Abstract
For further interpretation the apocarotenoid biosynthetic pathway in saffron, CsBCH1-z —a novel β carotene hydroxylase gene was isolated from stigma, and was cloned, which the full length of encoding region was 906 bps, contained 301 amino acid residues, and the corresponding to the genomic sequence of 1261bp, five exons separated by 4 introns. Compared with the amino acid sequence of the known CsBCH1 gene, there were two amino acids deletion and one amino acid changed from isoleucine to leucine caused by 6 bases deletion in the N- terminal, and another amino acid changed from threonine to alanine which was resulted in one SNP. It predicted CsBCH1-z protein contained 3 distinct transmembrane domains, C terminal contained two ‘HXXHH’ domain, was a typical fatty acid hydroxylase. As the key precursor of crocin synthesis, the expression level of CsBCH1-z was higher in stigma > leaves > petal, was very low in stamen, while was invisible in corm. The gene cloning and bioinformatics analysis of the novel saffron beta carotene hydroxylase provided the theoretical basis of the saffron active components biosynthetic process and accumulation.
关键词
西红花;β-胡萝卜素羟酶;克隆;生物信息学分析
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Key words
Saffron; beta carotene hydroxylase; Cloning; Bioinformatics analysis
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