臺灣雲杉雙功能雙萜合成酶基因之選殖與序列分析

Levopimaradiene/abietadiene synthase (LAS)為目前少數具有class I/II兩種功能的雙功能雙萜合成酶，其產物包括abietadiene、levopimaradiene 、neoabietadiene及palustradiene等存在於針葉樹樹脂中的雙萜類。本研究自資料庫中收集已發表之針葉樹LAS基因序列，並且在具有高度保守性的序列中設計一對退化性引子，對臺灣雲杉（Piceamorrisonicola Hayata）進行LAS基因的選殖，並配合RACE（Rapid Amplification of cDNA End）技術，成功獲得四個臺灣雲杉LAS基因部分片段。由分析比對的結果顯示，LAS為序列組成非常保守的基因，臺灣雲杉所包含的LAS基因片段與其他針葉樹種之基因相似度很高。此外，臺灣雲杉具有多個非常相似的LAS基因，如能了解其功能與作用差異，可有助於對臺灣雲杉在演化發展、分子標記與生化分類上的了解。

關鍵字

雙萜合成酶；基因選殖；臺灣雲杉

並列摘要

Levopimaradiene/abietadiene synthase (LAS) is not only a diterpene synthase that plays an important role in the synthesis of resin, it is also a rare class I/Ⅱ bifunctional diterpene synthase. This kind of enzyme produces multi-products such as levopimaradiene, abietadiene, neoabietadiene, and palustradiene. To investigate the sequence of LAS in Taiwan spruce (Picea morrisonicola Hayata), we used a pair of degenerate primers that were designed based on the conserved region of LAS genes from several conifer plants by PCR. Coupling with RACE (Rapid Amplification of cDNA End), we obtained four homologous genes from Taiwan spruce. The cloned genes were sequenced and their amino acid sequences predicted. According to the alignment data of nucleotides and amino acid sequences, there was a high similarity among the LAS genes from Taiwan spruce and the other conifer plants; therefore, the LAS gene was a highly conservative gene. Moreover, the study found several LAS genes in Taiwan spruce. They would provide helpful information about evolutionary development, molecular marker and phylogenetic relationship for Taiwan spruce once the differences among these LAS genes were comprehended.

並列關鍵字

Diterpene synthase ； Gene cloning ； Picea morrisonicola Hayata

國際替代計量

臺灣雲杉雙功能雙萜合成酶基因之選殖與序列分析

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