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首页> 外文期刊>Journal of genetics >Chalcone synthase genes from milk thistle (Silybum marianum): isolation and expression analysis
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Chalcone synthase genes from milk thistle (Silybum marianum): isolation and expression analysis

机译：乳蓟查尔酮合酶基因：分离和表达分析

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摘要

Silymarin is a flavonoid compound derived from milk thistle (Silybum marianum) seeds which has several pharmacological applications. Chalcone synthase (CHS) is a key enzyme in the biosynthesis of flavonoids; thereby, the identification of CHS encoding genes in milk thistle plant can be of great importance. In the current research, fragments of CHS genes were amplified using degenerate primers based on the conserved parts of Asteraceae CHS genes, and then cloned and sequenced. Analysis of the resultant nucleotide and deduced amino acid sequences led to the identification of two different members of CHS gene family, SmCHS1 and SmCHS2. Third member, full-length cDNA (SmCHS3) was isolated by rapid amplification of cDNA ends (RACE), whose open reading frame contained 1239 bp including exon 1 (190 bp) and exon 2 (1049 bp), encoding 63 and 349 amino acids, respectively. In silico analysis of SmCHS3 sequence contains all the conserved CHS sites and shares high homology with CHS proteins from other plants. Real-time PCR analysis indicated that SmCHS1 and SmCHS3 had the highest transcript level in petals in the early flowering stage and in the stem of five upper leaves, followed by five upper leaves in the mid-flowering stage which are most probably involved in anthocyanin and silymarin biosynthesis.
机译：水飞蓟素是从水飞蓟（Silybum marianum）种子衍生的类黄酮化合物，具有多种药理应用。查尔酮合酶（CHS）是类黄酮生物合成中的关键酶。因此，鉴定水飞蓟植物中CHS编码基因非常重要。在当前的研究中，使用基于菊科CHS基因保守部分的简并引物扩增CHS基因的片段，然后进行克隆和测序。分析所得的核苷酸和推导的氨基酸序列导致鉴定出CHS基因家族的两个不同成员，SmCHS1和SmCHS2。通过快速扩增cDNA末端（RACE）分离出第三成员全长cDNA（SmCHS3），其开放阅读框包含1239 bp，包括外显子1（190 bp）和外显子2（1049 bp），编码63和349个氨基酸， 分别。在计算机上分析SmCHS3序列包含所有保守的CHS位点，并与其他植物的CHS蛋白具有高度同源性。实时PCR分析表明，SmCHS1和SmCHS3在花的早期和花瓣的茎中有最高的转录本水平，在上面的五片叶子的茎中，随后在开花中的五片上面的叶子中，最可能与花色苷和水飞蓟素的生物合成。

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