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白木香乙酰乙酰基辅酶A硫解酶基因(AsAACT)的克隆与表达分析

Keywords: 白木香,AACT基因,MVA途径,序列分析,表达分析

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Abstract:

目的:对国产沉香的基原植物白木香Aquilariasinensis(Lour.)Gilg乙酰辅酶A酰基转移酶(acetyl-CoAC-acetyltransferase,AACT)基因AsAACT全长进行克隆并展开生物信息学分析和表达分析,为解析沉香萜类次生代谢产物的生物合成机制奠定基础。方法:根据获得的白木香转录组数据库AACT部分转录本序列设计引物,采用RT-PCR及RACE技术,以白木香茎cDNA为模板,克隆获得AsAACT全长,进行生物信息学分析;采用荧光定量PCR,以GADPH为内参,分析白木香愈伤组织受不同伤害胁迫AsAACT的表达模式。结果:AsAACT开放阅读框(openingreadingframe,ORF)为1236bp,编码411个氨基酸残基,酶命名为AsAACT;白木香愈伤受物理伤害(切割)后表达量没有明显变化,但受化学伤害(MeJA)后4h表达量升高了5.5倍,说明该基因对MeJA诱导的化学伤害较敏感,且能够在早期响应伤害胁迫。结论:通过AsAACT基因的全长cDNA克隆和表达特性分析,为后续深入研究其在沉香倍半萜合成途径的功能奠定基础。

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