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灵芝漆酶基因转录Cu2+诱导特性及其启动子的克隆与序列分析

DOI: 10.7685/j.issn.1000-2030.2009.01.008, PP. 36-40

Keywords: 灵芝,漆酶,半定量RT-PCR,self-formedadaptorPCR,转录调控

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Abstract:

以Cu2+为诱导物检测了灵芝漆酶同工酶基因的转录特性,并对其启动子进行了克隆及序列分析。研究发现灵芝漆酶基因在Cu2+的作用下表达量出现明显差异,其中以在发酵液中添加3.0mmol.L-1Cu2+、诱导时间为6d时,漆酶基因的mRNA表达量最高。根据GenBank中已报道的灵芝漆酶基因的序列信息,经PCR扩增获得了灵芝漆酶5′端长879bp的基因特异序列,进而通过self-formedadaptorPCR(SEFA-PCR)方法,扩增得到灵芝漆酶基因起始密码子上游长832bp的启动子序列。分析表明,该启动子区域除分布有TATA-box、CAAT-box及GC-box等基本的转录起始元件外,还存在多个潜在的顺式作用元件序列位点,包括4个MRE元件、4个STRE元件、11个HSE元件和5个氮因子结合位点等。

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