Title: Establishment of a normalized full-length cDNA library and bioinformatic analysis of ESTs of Miichthys miiuy
Abstract: To investigate molecular-assisted breeding in Miichthys miiuy,a normalized full-length cDNA library was established using the duplex-specific nuclease(DSN) normalization method combined with switching mechanism at 5' end of RNA transcript(SMART) technique.The titer of un-amplified cDNA library was about 1.6×106 CFU·mL-1;the cDNA library contained 7.5×106 independent clones with DNA inserts of 1-3 kb.A total of 5952 clones were randomly selected to be sequenced and 5053 expressed sequence tags(EST) were obtained,and 4609 high quality EST sequences were generated.After processing,a total of 3221 unigenes comprising 656 contigs and 2565 singlets were obtained,and the redundancy ratio was about 30.11%.The average reading length was 550 bp,and reading length of most sequences was ranged from 500 to 599 bp and the GC contents of most sequences was ranged from 40 to 60 percentages.A total of 1397 functional known genes were categorized by gene ontology.Many putative immune genes were identified that encoded major histocompatibility complex,cytokines and cytokine receptors,adhesive proteins,stress proteins,transcription factors for immune response,immunoglobulin and coagulation factors.The results above provide both a detailed annotation of immune genes and a collection of novel transcripts for this important species.
Publication Year: 2011
Publication Date: 2011-11-25
Language: en
Type: article
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