| Chicken-claw-shaped Polyporus umbellatus is a unique source in Changbai Mountains area,which has experienced three period of life history from mycelium,sclerotium to fruit body.The expressed sequence tags(EST)of sclerotium,developed for further study of growth,metabolic regulation,resources utilization and protection of rare medicinal fungi,has provided a theoretical foundation.The raw material was chicken-claw-shaped polyporus umbellatus sclerotiam which collected from the Tiannan township XieFang Village of Jiaohe city in Jilin province.The yield of total RNA was 6.24 ug extracted by the improving Trizol method.The cDNA library was constructed by SMARTTM cDNA Library Construction Kit.The titer and recombination rate of the unamplified library were 3.4 x 106 pfu/mL and 93.68%.The titer and capacity of amplified library were 1.304 x 1010 pfu/mL and 3.13 x 1012 pfu.A total of 500 clones randomly selected from the cDNA library producted 452 high quality ESTs and,from which 369 unigenes were annotated.The redundancy of cDNA library was 23%.In arabidopsis thaliana function classification criteria,78 known function unigenes by BLAST x was divided into 9 classes,respectively,metabolism,energy,transcription,protein synthesis,cell structure,transport,signal transduction,defense and disease and others.Only 183 unigenes got one or more than one GO term,which cell component consisted of seven parts,molecular function included five parts,biochemical processes included 16 parts.Twelve full ESTs sequence containing the open reading frame and conserved domain,has forecasted the encoded proteins and biological information.As a result of successfully extracting the total RNA and constructing a high quality cDNA library.EST analysis of formation and development mechanism of chicken-claw-shaped polyporus umbellatus sclerotium,and for specificity clone of the functional gene,has laid a theoretical foundation. |
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