Soden D M, Dobson A D W
National Food Biotechnology Centre, University College Cork, National University of Ireland, Cork, Ireland.
J Appl Microbiol. 2003;95(3):553-62. doi: 10.1046/j.1365-2672.2003.02012.x.
To determine the regulation of laccase isozyme gene transcription in Pleurotus sajor-caju in response to different aromatic inducers and physiological parameters.
The promoter regions for each of four different laccase isozymes were cloned from P. sajor-caju, using amplified flanking region-PCR (AFR-PCR). Sequences stretching 724, 214, 840 and 1740 bp upstream from the predicted start codons for lac1, lac2, lac3 and lac4, respectively, were cloned in each case and analysed for the presence of putative transcriptional response elements. A number of putative response elements including metal response elements, xenobiotic response elements and antioxidant response elements appear to be present. In addition putative consensus sequences such as those for the binding of AP1, AP2, creA and NIT2 transcription factors, which are involved in nitrogen and carbon regulation in different fungi, are also present in the promoter regions of some of the isozymes.
These elements may be involved in the transcriptional regulation of laccase gene expression in P. sajor-caju.
The presence of a number of putative transcriptional response elements in the promoter regions of different isozyme genes indicates a potential role for these sites in regulating laccase gene transcription in P. sajor-caju. In addition this work demonstrates the potential usefulness of AFR-PCR as a technique to clone fungal DNA sequences located upstream from known sequences.
确定凤尾菇漆酶同工酶基因转录对不同芳香族诱导剂和生理参数的响应调控。
采用扩增侧翼区域PCR（AFR-PCR）技术，从凤尾菇中克隆了四种不同漆酶同工酶各自的启动子区域。分别克隆了位于lac1、lac2、lac3和lac4预测起始密码子上游724、214、840和1740 bp的序列，并分析了假定的转录反应元件的存在情况。似乎存在许多假定的反应元件，包括金属反应元件、外源性反应元件和抗氧化反应元件。此外，一些同工酶的启动子区域还存在假定的共有序列，如参与不同真菌氮和碳调控的AP1、AP2、creA和NIT2转录因子结合的共有序列。
这些元件可能参与凤尾菇漆酶基因表达的转录调控。
不同同工酶基因启动子区域存在许多假定的转录反应元件，表明这些位点在调控凤尾菇漆酶基因转录中具有潜在作用。此外，这项工作证明了AFR-PCR作为一种克隆已知序列上游真菌DNA序列的技术的潜在实用性。
