Development And Application Of Analysis Platform For High-Throughput ESTs From CDNA Library Of Compatible Interaction Between Wheat And Puccinia Striiformis F. Sp. Tritici

To analyze large-scale ESTs (Expressed Sequence Tags, ESTs) sequences obtained from the cDNA libraries of interaction between wheat and Puccinia striiformis f. sp. tritici can not only get the gene expression profiles during interaction between host plant and pathogen, but also obtain the genes in relation to infection or resistance. All of the analysis to ESTs plays an important role in revealing the molecular mechanism of interaction between wheat and P. striiformis and cultivating new resistant wheat varieties.Large amounts of ESTs were obtained from cDNA library of interaction between wheat and P. striiformis constructed in our laboratory. To analyze the data manually is tough, leading to low efficiency and poor accuracy, especially the homological analysis on the Internet. Therefore, the local ESTs analysis platform and related local databases were developed which can analyze the large-scale ESTs with local computer quickly and efficiently.1 A local high-throughput ESTs analysis platform was developed. It consisted of sequence pre-operating system, ESTs homological analysis system and ESTs function annotation system, which can accomplish the process from inputting chromatography file to outputting results of function annotation about large-scale ESTs data. The practice of analyzing large-scale ESTs from cDNA library of compatible interaction between wheat and P. striiformis showed that the platform is efficient and easy-operating.2 Totals of 6002 ESTs sequences from the cDNA library of compatible interaction between wheat and P. striiformis were clustered using this platform, resulting in 2746 uniseq, which contained 610 contigs and 2136 singlets. Function annotation was performed on the 2746 uniseq based on GO (Gene Ontology), KO (KEGG Ontology) and protein databases of ath and sce (local databases) respectively. Gene expression profiles and information about pathway were obtained and it is beneficial to the next research on gene function furthermore.The local high-throughput ESTs analysis platform is an integrated bioinformatics system, which can do varieties bioinformatics analysis to biological data acquired in our research. The special databases (eg, cogeme fungal ESTs database) can provide a better service to the research on interaction between host and pathogen in our laboratory.