原晓龙;赵能;华梅;陈剑;杨宇明;王娟;王毅;

• 1:云南省林业科学院云南省森林植物培育与开发利用重点实验室国家林业局云南珍稀濒特森林植物保护和繁育重点实验室
• 2:西南林业大学林学院

摘要(Abstract)：

为了解牛樟芝三萜类生物合成、调控机理,以多孔菌科真菌茯苓(Wolfiporia cocos,AFR13032.1)的SQS基因为模板对牛樟芝基因组数据进行本地Blast,分离获得牛樟芝鲨烯合酶(AcSQS)基因.以该序列为模板设计特异引物Ac SQSF0和Ac SQSR0,通过PCR扩增得到Ac SQS c DNA全长,并对其进行生物信息学分析,检测不同碳氮源添加物的培养基上该基因的表达水平.结果显示:克隆得到的基因为牛樟芝鲨烯合酶(Ac SQS),该基因含4个外显子,3个内含子,外显子拼接总长1 803 bp,编码600个氨基酸;其蛋白序列的1¹7位为信号肽位点,具两段跨膜结构,可能定位于内质网;位于该蛋白18917位为信号肽位点,具两段跨膜结构,可能定位于内质网;位于该蛋白189²04位的CHYVAGLVGEGLTRL和225204位的CHYVAGLVGEGLTRL和225²38位的MGLMLQKTNIIRDY的保守基序为鲨烯合酶识别位点,DTIEDD和D(Y/F)RED为FPP绑定位点;蛋白网络分析显示其位于三萜类化合物的代谢网络中;聚类分析显示Ac SQS蛋白和茯苓、硫磺多孔菌、拟蜡菌、灵芝、云芝等的SQS蛋白聚为一支;不同碳氮源添加物培养基上的表达分析显示:果糖诱导该基因表达的能力最强,而不同氮源添加物诱导该基因表达的能力无明显差异.

关键词(KeyWords)： 牛樟芝;鲨烯合酶;基因克隆;表达分析

Abstract:

Keywords:

基金项目(Foundation): 对外科技合作计划项目(2015IA004);; 国家自然科学基金青年项目(31400488);; 云南省应用基础研究计划面上项目(2016FB055)

作者(Authors): 原晓龙;赵能;华梅;陈剑;杨宇明;王娟;王毅;

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