Kim Ok Tae, Kim Min Young, Huh Sun Mi, Bai Dong Gyu, Ahn Jun Cheul, Hwang Baik
Department of Biology, Chonnam National University, Gwangju, 500-757, South Korea.
Plant Cell Rep. 2005 Jul;24(5):304-11. doi: 10.1007/s00299-005-0927-y. Epub 2005 Apr 15.
A homology-based PCR method was used to clone a cDNA encoding oxidosqualene cyclase from Centella asiatica, which produces a large quantity of triterpene saponins such as asiaticoside and madecassoside. Sequence analysis of one clone found sequences related to beta-amyrin synthase. An open reading frame in the full-length clone was named CabAS (Centella asiatica putative beta-amyrin synthase). On the basis of amino acid sequence, CabAS appears to be an enzyme (beta-amyrin synthase) that synthesizes beta-amyrin. Southern analysis showed that the C. asiatica genome contains one copy of the CabAS gene. Northern blot analysis demonstrated that the CabAS gene is expressed in leaves with no detectable transcript in other plant tissues, consistent with the organ-specific accumulation of the asiaticoside. Up-regulation of expression of CabAS by methyl jasmonate in leaves was also demonstrated.
采用基于同源性的聚合酶链反应(PCR)方法,从积雪草中克隆了一个编码氧化鲨烯环化酶的互补脱氧核糖核酸(cDNA),积雪草能产生大量三萜皂苷,如积雪草苷和羟基积雪草苷。对一个克隆进行序列分析,发现了与β-香树脂醇合酶相关的序列。全长克隆中的一个开放阅读框被命名为CabAS(积雪草假定β-香树脂醇合酶)。根据氨基酸序列,CabAS似乎是一种合成β-香树脂醇的酶(β-香树脂醇合酶)。Southern分析表明,积雪草基因组包含一个CabAS基因拷贝。Northern印迹分析表明,CabAS基因在叶片中表达,在其他植物组织中未检测到转录本,这与积雪草苷的器官特异性积累一致。还证明了茉莉酸甲酯可上调叶片中CabAS的表达。