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查尔酮合成酶(chalcone synthase,CHS)是黄酮类物质合成的第一关键酶。为了获得CHS基因,我们在ACGM标记所扩增的PCR片段内设计锚定引物,并结合RACE技术首次克隆了柳杉属3种植物的CHS基因,包括柳杉(Cryptomeria japonica var.sinensis,CjsCHS)、短茸柳杉(C.japonicacv.Araucarioides,CjaCHS)和日本柳杉(C.japonica,CjCHS),并对其进行分析。结果表明:3个基因长都为1 176bp,编码391个氨基酸,它们都含有CHS高度保守活性位点以及CHS标签序列GFGPG,其编码蛋白的相似度达99%,与其他植物相似度在79%以上,表明CHS基因在进化上具有相对保守性。
Chalcone synthase (CHS) is the first key enzyme in flavonoid synthesis. In order to obtain the CHS gene, we designed the anchor primers in the PCR fragments amplified by ACGM and cloned the CHS genes of three species of Cryptomeria including Cryptomeria japonica var. Sinensis (CjsCHS) , C.japonicacv. Araucarioides (CjaCHS) and C. japonica (CjCHS) were analyzed and analyzed. The results showed that all three genes were 1 176 bp in length and encoded 391 amino acids. All of them contained the highly conserved CHS active site and the CHG tagged sequence GFGPG. The similarity of the encoded proteins was 99% and the similarity with other plants was 79% The above shows that CHS gene is relatively conservative in evolution.