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当前位置: 首页 > 文章 > 北柴胡鲨烯合酶基因及其编码区cDNA克隆与序列分析 园艺学报 2010,37 (2) 283-290
Position: Home > Articles > Cloning and Sequence Analysis of Squalene Synthase Gene and cDNA in Bupleurum chinense DC. Acta Horticulturae Sinica 2010,37 (2) 283-290

北柴胡鲨烯合酶基因及其编码区cDNA克隆与序列分析

作  者：
隋春;魏建和;战晴晴;杨成民
单  位：
中国医学科学院
关键词：
北柴胡;柴胡皂苷;鲨烯合酶基因(SS);RT-PCR;PCR
摘  要：
鲨烯合酶(EC.2.5.1.21,squalene synthase,SS)是植物甾醇和三萜化合物生物合成途径中的关键酶。为研究鲨烯合酶在柴胡皂苷生物合成中的作用,以北柴胡(Bupleurum chinense DC.)不定根为试材,采用Trizol法提取总RNA,利用RT-PCR方法从北柴胡中扩增出了鲨烯合酶cDNA特异片段,并进行了克隆、测序。测序结果显示得到了两个序列不同的cDNA(BcSS1和BcSS2),分别为1245bp和1248bp,分别编码414及415个氨基酸。BcSS1和BcSS2间的核苷酸和氨基酸一致性分别为98%和96%。NCBI Blastx结果显示与三岛柴胡和三七SS氨基酸序列相似性最高,BcSS1的一致性分别为99%和90%,BcSS2的分别为97%和87%。在此基础上,利用PCR技术从北柴胡不定根中扩增得到了一个SS基因克隆,长5880bp,包含12个内含子。
译  名：
Cloning and Sequence Analysis of Squalene Synthase Gene and cDNA in Bupleurum chinense DC.
作  者：
SUI Chun1,WEI Jian-he1,ZHAN Qing-qing1,2,and YANG Cheng-min1 (1Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100193,China;2College of Life Science,Northeast Forestry University,Harbin 150040,China)
关键词：
Bupleurum chinense DC.;saikosaponin;squalene synthase gene(SS);RT-PCR;PCR
摘  要：
Squalene synthase(EC.2.5.1.21,SS)is an important key enzyme in biosynthesis pathway of plant sterols and terpenoids.In order to learn the role of SS in the biosynthesis of saikosaponin in Bupleurum chinense DC.,SS cDNA was cloned with total RNA extracted from adventitious roots of B.chinense using Trizol method.Specific fragments were amplified by RT-PCR and then were cloned and sequenced.Sequencing results showed two different cDNA fragments(BcSS1 and BcSS2)with 1 245 bp and 1 248 bp long were obtained which encodes proteins with 414 and 415 amino acids,respectively.The identities of nucleotides and amino acids between BcSS1 and BcSS2 were 98% and 96%.NCBI Blastx search results showed BcSS1 and BcSS2 had the highest amino acid similarity to the corresponding proteins from B.falcatum L.and Panax notoginseng F.H.Chen.The identities of BcSS1 with the two proteins were 99 % and 90% and those of BcSS2 were 97% and 87%.SS gene with 5 880 bp containing 12 introns was then amplified by PCR with genomic DNA extracted from adventitious roots of B.chinense using CTAB method.

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