PCR-MEDIATED ANALYSIS OF LIGNOCELLULOLYTIC GENE-TRANSCRIPTION BY PHANEROCHAETE-CHRYSOSPORIUM - SUBSTRATE-DEPENDENT DIFFERENTIAL EXPRESSION WITHIN GENE FAMILIES
PCR-MEDIATED ANALYSIS OF LIGNOCELLULOLYTIC GENE-TRANSCRIPTION BY PHANEROCHAETE-CHRYSOSPORIUM - SUBSTRATE-DEPENDENT DIFFERENTIAL EXPRESSION WITHIN GENE FAMILIES
复制标题
DOI：
10.1128/aem.61.6.2358-2364.1995
复制
发表时间：
1995-06-01
影响因子：
4.4
通讯作者：
SIMS, PFG
中科院分区：
文献类型：
--
作者：
BRODA, P;BIRCH, PRJ;SIMS, PFG
我们比较了在四种不同碳源上生长的黄孢平革菌ME446上清酶活性（木质素过氧化物酶、锰过氧化物酶和纤维素酶）和基因表达（LIG、mnp和cbhI基因家族以及独特的cbhII基因）的表观动力学：代表天然底物木质纤维素的磨粒秸秆；Avicel为结晶纤维素；高浓度和低浓度的葡萄糖，在所有情况下都有限制氮。利用对特定基因特异性引物的pcr技术表明，家族之间和家族内部存在差异表达。有许多mRNA物种仅在一天内存在的实例，这意味着在mRNA水平上对木质纤维素降解有严格的调节。细胞外酶活性及mnp和cbh基因表达模式相似，而当细胞外上清中未观察到相应酶活性时，可检测到LIG基因表达。在这些条件下产生的酶可能被菌丝体隔离，并且可能具有重要的功能。另一个惊人的结果是纤维素，以Avicel的形式，引发了三个LIG基因的表达，而这些基因在与其他碳源相同的条件下没有表达。
We compare the kinetics of appearance of supernatant enzyme activities (lignin peroxidase, manganese peroxidase, and cellulase) and gene expression (LIG, mnp, and cbhI gene families and the unique cbhII gene) in Phanerochaete chrysosporium ME446 when grown on four different carbon sources: hall-milled straw, representing the natural substrate lignocellulose; Avicel as a crystalline cellulose; and high and low concentrations of glucose, in all cases with limiting nitrogen. PCR-based technology utilizing pairs of primers specific for particular genes showed that there is differential expression between and within the families. There were a number of instances of mRNA species being present only on a single day, implying tight regulation of lignocellulose degradation at the mRNA level. The patterns of extracellular enzyme activities and mnp and cbh gene expression are similar whereas LIG gene expression can be detected when no corresponding enzyme activity is observed in the extracellular supernatant. The enzyme produced under these conditions is presumably sequestered by the mycelium and is likely to be functionally significant. Another striking result is that cellulose, in the form of Avicel, elicits the expression of three LIG genes for which there is no expression under the same conditions with the other carbon sources.