Construction And Part Of Bioinformatics Analysis Of Cdna Libraryof Sclerotium Of Medicinal Funguspolyporus Umbellatus

Polyporus umbellatus(Pers.)Fr.is a medicinal fungus that has diuretic and edematous effects;it has three stages of growth:sclerotia,mycelia,and fruit bodies,of which sclerotia are medicinal.In part,this thesis studies the ESTs of the porcine sclerotia sclerotiorum and provides a theoretical basis for the further study of the formation mechanism of the sclerotia sclerotiorum and the analysis of the genomic differences in species.Using the three-year-old sclerotia from Foping County,Shaanxi Province as raw material,total RNA was extracted by the EASY spin plant RNA kit.Electrophoretic detection of bands is clear and the OD260/280 was between 2.0 and 2.1.Transcription requirements:A cDNA library of S.solani was successfully constructed using the SMART method.The titer of the original library was 6.55×106 pfu/ml,the recombination rate was 94.26%,and the length of the inserted genes was greater than 100bp.The titer of the amplified library was 1.821×1010 pfu/ml,in line with a better quality library standard.500 randomly selected single clones were sequenced in one direction,and a total of 371 high-quality sequences were obtained with an average length of 383 bp.After clustering and assemble,a total of 46 contigs and 244 singletons containing 127 ESTs were obtained,resulting in a total of 290 unigenes,the redundancy of the library was 21.83%;The reference functional list of plant genes classifies the known functional sequences into 11 categories:protein synthesis,disease/defense,metabolism,signal transduction,energy,transport,cell growth/division,transcription,cell structure,intracellular pathway and secondary metabolism.Only 160 sequences obtained one or more GO functional gene annotations;9 functions were annotated as cellular components;15 functions were annotated as biological processes;3 functions were annotated as molecular functions.The resulting sequence was submitted to the KEGG database for comparison.A total of 116 genes obtained K number,of which 98 genes were brought into the metabolic pathway,involving a total of 153 metabolic pathways;of which the most involved pathways are metabolism,A total of 22 genes were involved;10 sequences were selected to predict the open reading frame and the conserved domain of the protein.The closest known protein of the gene was found by blast and the properties and conformation of known proteins were analyzed.