【作者】 欧阳翔； 吴婧； 丁一新； 李玉祥； 赵明文；

【Author】 Ou-yang Xiang,Wu jing,Ding Yi-xin,Li Yu-xiang,Zhao Ming-wen* (College of Life Sciences,Nanjing Agricultural University;Key Lab of Microbiological Engineering of Agricultural Environment,Ministry of Agriculture,Nanjing 210095)

【机构】 南京农业大学生命科学学院农业部农业环境微生物工程重点开放实验室；

【摘要】 本文以Cu²⁺为诱导物,检测灵芝漆酶同工酶基因的转录特性。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,其中以发酵液中添加3.0 mmol·L^-1Cu²⁺、诱导时间为6天时,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,获得了韩国灵芝漆酶5′端的基因特异序列,进而通过self-formed adaptor PCR（简称SEFA-PCR）的方法,扩增得到灵芝漆酶基因起始密码子上游长802 bp的启动子序列。分析表明,该启动子区域上除分布有TATA-box、CAAT-box以及GC-Box等基本的转录起始元件外,还存在有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HSEs元件和5个氮因子结合位点等。这些结果表明,不同的外源诱导物可以调节灵芝漆酶基因的表达。更多还原

【Abstract】 Copper was taken as an inducer to investigate the transcription characteristics of the laccase isozyme gene from Ganoderma Lucidum.The expression of the laccase gene in the G Lucidum was significantly different by the effects of copper in our research.The expression of mRNA of the laccase gene was the highest after treated by adding 3.0 mmol?L^-1 copper into the liquid medium for 6 days.According to the sequence of the laccase gene of the G lucidum from GenBank,the 5’ terminal sequence of the laccase gene of G lucidum HG was obtained.Then a 802 bp promoter sequence on the upstream of the initiation codon in laccase gene of the G Lucidum HG was amplified through a self-formed adaptor PCR（termed SEFA PCR） method.The promoter was analyzed using the Softberry software.The result showed that,besides some basic transcription initiation elements,such as TATA-box,CAAT-box and GC-Box,some other potential cis-acting element sequence sites were also existed,including four MREs,four STREs,eleven HSEs and five nitrogen factor binding sites,etc.These results proved that the expression of the laccase gene of the G Lucidum can be regulated by different exterior inducers.更多还原