| The sea area of Zhoushan Islands are rich in fishery resources and rich in fish species.In this study,transcriptome sequencing was carried out for 5 species of fish with high economic value in Zhoushan,including Conger myriaster,Konosirus punctatus,Liza affinis,Acanthopagrus schlegelii,Acanthogobius ommaturuswas.The problem of lack of transcriptome information in the above five species of fish was solved.The research results are as follows:The original data of the five species of fish tested in this study ranged from 40300 666(Liza affinis)to 55188198(Conger myriaster)after quality control.The data after quality control ranges from 6.05 G to 8.287 G,respectively.The transcripts were spliced by Trinity and the length of N50 ranged from 906 bp to 2791 bp and from 255 bp to 676 bp respectively.In addition to Tr EMBL database annotations for K.punctatus transcription group,gene functional annotations were completed after comparing seven databases: CDD,KOG/COG,NR,NT,Pfam and Swiss-prot.The maximum coverage rate was61.41%-86.51%.The protein coding sequence of each fish species was successfully predicted.The protein sequences of C.myriaster,K.punctatus,L.affinis,A.schlegelii,A.ommaturuswas were 29807,53053,25229,32934 and 24405 respectively.SSR prediction and primer design were carried out on the transcriptome data of each fish species,and many high-score SSR primers were designed.The analysis of gene expression in each fish species showed that there were significant differences in gene expression among different species,but there were similarities among them.The highest expression level in the transcriptome of C.myriaster and A.schlegelii is actin and related protein genes.The expression level of cytochrome c oxidase related protein in the C.myriaster,L.affinis,A.schlegelii is in the top five.The expression level of chymotrypsin precursors in K.punctatus larvae was relatively high.A.ommaturuswas highly expresses an antimicrobial peptide.The expression levels of each species ranged from high to low,ranking the top five:The top five genes in C.myriaster gene expression level are actin and related protein genes,calmodulin and related protein genes(EF-hand superfamily),cytochrome c oxidasesubunit ? and related protein genes,myosin heavy chain related genes and fructose-1,6-diphosphate aldolase related genes.The top five genes in K.punctatus gene expression level are trypsin-like serine protease related genes,chymotrypsinogen-like protein fragment-related genes,pre-splicing factor SPF27 related protein genes,actin and related protein genes,and chymotrypsinogen-related genes.The top five genes in L.affinis gene expression level are cytochrome c oxidase subunit I protein-related genes,cytochrome oxidase subunit ? and related proteins,nucleoside diphosphate(NDP)kinase,? hemoglobin B gene and fibrinogen ? chain protein-related genes.The top five genes in A.schlegelii gene expression level are skeletal muscle actin and related protein genes,albumin beta protein related genes,cardiac actin related genes,cytochrome c oxidase related genes,creatine kinase related genes.The top five genes in A.ommaturuswas gene expression level are Hepcidin-like protein gene,apolipoprotein A-? related gene,hypothetical protein,uncharacterized protein and intracellular protein transport-related gene.Based on the protein sequences obtained from transcriptome data,combined with six fish proteome sequences downloaded from NCBI database,single copy genes were extracted by Ortho MCL and phylogenetic tree was constructed.The C.myriaster and Acanthopterygii were divided into two distinct branches,of which Gobiidae were clustered into a very significant single branch;Mugiliformes L.affinis and Perciformes Stegastes partitus were clustered together;the remaining three Perciformes species were clustered into one. |
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