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查尔酮合酶(CHS)是类黄酮生物合成过程中的第一个关键酶,其在类黄酮的合成过程中发挥着非常重要的作用。本研究以日本蛇根草为研究材料,根据其转录组测序结果设计全长引物,通过RT-PCR方法成功克隆得到日本蛇根草查尔酮合酶基因完整的cDNA序列,并利用相关生物信息学软件对其进行分析。结果表明,该基因序列全长为1 170 bp,编码389个氨基酸,预测其蛋白分子量为42.783 kD,等电点为6.05,是亲水性蛋白质,不含有信号肽,很可能定位在细胞质中。同时,二级结构分析显示,日本蛇根草CHS的二级结构分别由180个α螺旋、84个延伸链、125个无规则卷曲组成,这与三维结构预测结果相一致。日本蛇根草CHS基因的克隆对于研究植物类黄酮的生物合成及其分子机制具有重要意义,同时也丰富了双子叶植物PKS基因家族的相关研究。
Chalcone synthase (CHS) is the first key enzyme in flavonoid biosynthesis and plays a very important role in the synthesis of flavonoids. In this study, the Japanese snakehead grass as a research material, according to its transcriptome sequencing results designed full-length primers, cloned by RT-PCR method cloned snake root grass chalcone synthase gene complete cDNA sequence, and use of relevant biological information Learn software to analyze it. The results showed that the full length of this gene was 1 170 bp encoding a polypeptide of 389 amino acids. The predicted protein molecular weight was 42.783 kD and the isoelectric point was 6.05. It was a hydrophilic protein with no signal peptide and was probably located in the cytoplasm. At the same time, secondary structure analysis showed that the secondary structure of CHS in Japanese snakehead grass was composed of 180 α-helices, 84 extended chains and 125 random curls, which was consistent with the predicted results of three-dimensional structure. The cloning of CHS gene of Japanese snakehead grass is of great significance for studying the biosynthesis of plant flavonoids and its molecular mechanism, and also enriches the research on the PKS gene family of dicotyledonous plants.