Isolation and identification of small species-specific secreted proteins of Marssonina brunnea
作者单位：Jiangsu Key Laboratory for Poplar Germplasm Enhancement and Variety ImprovementNanjing Forestry University
会议名称：《2013全国植物生物学大会》
会议日期：2013年
关 键 词：effector Consensus-Degenerate Hybrid Oligonucleotide Primer degenerate PCR
摘 要：Fungal plant pathogens produce proteinaceous effectors that can increase their pathogenicity. These fungal effectors are frequently small secreted proteins with limited phylogenetic *** a proteomics approach for analyzing the secretome is a promising strategy for the identification of effector proteins of phytopathogenic ***,the primary limitation of this strategy is the low identification rate of spectrometry-based proteomic methods that rely heavily on the accuracy and integrity of the reference sequence database. Marssonina brunnea is an important fungal pathogen of the Populus *** further our understanding of the pathogenesis of ***,we initiated a proteome-level study of the fungal *** de novo peptide sequencing by MS/MS,we obtained peptide sequences for 32 protein *** proteins were identified by sequence homology to conserved proteins in public databases using MS-driven *** identify additional protein spots,we combined a degenerate PCR method,based on the Consensus-Degenerate Hybrid Oligonucleotide Primer(CODEHOP) method,and a rapid amplification of cDNA ends method to clone the full-length cDNA fragments encoding the proteins identified in the gel. Using this method,we cloned the full-length cDNA fragments encoding 9 putative Lysm effector *** method provides an efficient approach to identification of species specific effector proteins of non-sequenced fungal organisms.