Departamento de Microbiologia, Universidade Federal de Viçosa, Viçosa, MG Brazil.
Genet Mol Biol. 2009 Apr;32(2):362-6. doi: 10.1590/S1415-47572009000200024. Epub 2009 Jun 1.
This report describes the cloning, sequence and expression analysis of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene of Moniliophthora perniciosa, the most important pathogen of cocoa in Brazil. Southern blot analysis revealed the presence of a single copy of the GAPDH gene in the M. perniciosa genome (MpGAPDH). The complete MpGAPDH coding sequence contained 1,461 bp with eight introns that were conserved in the GAPDH genes of other basidiomycete species. The cis-elements in the promoter region of the MpGAPDH gene were similar to those of other basidiomycetes. Likewise, the MpGAPDH gene encoded a putative 339 amino acid protein that shared significant sequence similarity with other GAPDH proteins in fungi, plants, and metazoans. Phylogenetic analyses clustered the MPGAPDH protein with other homobasidiomycete fungi of the family Tricholomataceae. Expression analysis of the MpGAPDH gene by real-time PCR showed that this gene was more expressed (~1.3X) in the saprotrophic stage of this hemibiotrophic plant pathogen than in the biotrophic stage when grown in cacao extracts.
本报告描述了巴西可可最重要病原体蜜环菌甘油醛-3-磷酸脱氢酶（GAPDH）基因的克隆、序列和表达分析。Southern blot 分析显示，M. perniciosa 基因组（MpGAPDH）中存在单个 GAPDH 基因拷贝。MpGAPDH 完整编码序列包含 1461bp，具有 8 个内含子，与其他担子菌属物种的 GAPDH 基因保守。MpGAPDH 基因启动子区的顺式元件与其他担子菌属相似。同样，MpGAPDH 基因编码一个假定的 339 个氨基酸蛋白，与真菌、植物和后生动物中的其他 GAPDH 蛋白具有显著的序列相似性。系统发育分析将 MPGAPDH 蛋白与伞菌科的其他同担子菌属真菌聚类在一起。实时 PCR 表达分析显示，与在可可提取物中生长的生物营养阶段相比，该基因在这种半腐生植物病原体的腐生阶段的表达更为显著（~1.3X）。
