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有参转录组学习七:基因功能注释及富集分析

有参转录组学习七:基因功能注释及富集分析

作者: 颤抖吧__小虫子 | 来源:发表于2019-05-18 19:09 被阅读0次

    Author: ligc

    Date: 19/5/18

    library(AnnotationHub)
    library(clusterProfiler)
    ##安装旧版本的包
    packageurl <- "https://cran.r-project.org/bin/windows/contrib/3.5/gridGraphics_0.3-0.zip"
    install.packages(packageurl,repos=NULL, type="source")
    AH <- AnnotationHub()
    AH
    AnnotationHub with 47474 records
    # snapshotDate(): 2018-10-24 
    # $dataprovider: BroadInstitute, Ensembl, UCSC, ftp://ftp.ncbi....
    # $species: Homo sapiens, Mus musculus, Drosophila melanogaster...
    # $rdataclass: GRanges, BigWigFile, FaFile, TwoBitFile, OrgDb, ...
    # additional mcols(): taxonomyid, genome, description,
    #   coordinate_1_based, maintainer, rdatadateadded,
    #   preparerclass, tags, rdatapath, sourceurl, sourcetype 
    # retrieve records with, e.g., 'object[["AH2"]]' 
    
                title                                               
      AH2     | Ailuropoda_melanoleuca.ailMel1.69.dna.toplevel.fa   
      AH3     | Ailuropoda_melanoleuca.ailMel1.69.dna_rm.toplevel.fa
      AH4     | Ailuropoda_melanoleuca.ailMel1.69.dna_sm.toplevel.fa
      AH5     | Ailuropoda_melanoleuca.ailMel1.69.ncrna.fa          
      AH6     | Ailuropoda_melanoleuca.ailMel1.69.pep.all.fa        
      ...       ...                                                 
      AH67895 | org.Vibrio_vulnificus.eg.sqlite                     
      AH67896 | org.Achromobacter_group_B.eg.sqlite                 
      AH67897 | org.Achromobacter_group_E.eg.sqlite                 
      AH67898 | org.Pannonibacter_phragmitetus.eg.sqlite            
      AH67899 | org.Salinispora_arenicola_CNS-205.eg.sqlite 
    
    数据来源
    unique(AH$dataprovider)
    [1] "Ensembl"                              
     [2] "UCSC"                                 
     [3] "RefNet"                               
     [4] "Inparanoid8"                          
     [5] "NHLBI"                                
     [6] "ChEA"                                 
     [7] "Pazar"                                
     [8] "NIH Pathway Interaction Database"     
     [9] "Haemcode"                             
    [10] "BroadInstitute"                       
    [11] "PRIDE"                                
    [12] "Gencode"                              
    [13] "CRIBI"                                
    [14] "Genoscope"                            
    [15] "MISO, VAST-TOOLS, UCSC"               
    [16] "UWashington"                          
    [17] "Stanford"                             
    [18] "dbSNP"                                
    [19] "BioMart"                              
    [20] "GeneOntology"                         
    [21] "KEGG"                                 
    [22] "URGI"                                 
    [23] "EMBL-EBI"                             
    [24] "MicrosporidiaDB"                      
    [25] "FungiDB"                              
    [26] "TriTrypDB"                            
    [27] "ToxoDB"                               
    [28] "AmoebaDB"                             
    [29] "PlasmoDB"                             
    [30] "PiroplasmaDB"                         
    [31] "CryptoDB"                             
    [32] "TrichDB"                              
    [33] "GiardiaDB"                            
    [34] "ftp://ftp.ncbi.nlm.nih.gov/gene/DATA/"
    
    head(unique(AH$species))
    [1] "Ailuropoda melanoleuca" "Anolis carolinensis"    "Bos taurus"            
    [4] "Caenorhabditis elegans" "Callithrix jacchus"     "Canis familiaris" 
    
    #查找具体的物种信息
    grs <- query(AH,"Mus musculus ")
    #AH[AH$species == 'Arabidopsis thaliana' & AH$rdataclass == 'OrgDb']
    #subset(AH, species == 'Arabidopsis thaliana' & rdataclass == 'OrgDb')
    #图形化
    display(AH)
    unique(AH$rdataclass)
    ##下载
    mus <- AH[["AH53222"]]
    class(mus)
    columns(mus)
    keytypes(mus)
    head(keys(mus,keytype = "SYMBOL"))
    select(mus,keys = "0610006L08Rik",columns = c("GENEID","GO"),keytype = "SYMBOL")
    mus.sample <- sample(keys(mus,keytype = "ENTREZID"),200)
    test <- enrichGO(gene          = mus.sample,
                     OrgDb         = mus,
                     keyType       = "ENTREZID",
                     pAdjustMethod = "none",
                     pvalueCutoff  = 0.1,
                     qvalueCutoff  = 0.2)
    mapIds(mus, keys = mus.sample, column = c("TXID"), keytype = "ENTREZID")
    ### keys是原始的ID,columns是转换之后的ID,keytype是要指定的原始ID类型
    BiocManager::install("org.Mm.eg.db")
    library(org.Mm.eg.db)
    keytypes(org.Mm.eg.db)
    columns(org.Mm.eg.db)
    select(org.Mm.eg.db,keys = "ENSMUSG00000001123",columns = c("SYMBOL","GENENAME","GO"),keytype = "ENSEMBL")
    mouse_DEG_list <- read.table(file = "mouse_DEG_list",header = T,row.names = 1)
    mouse_DEG_list_name <- row.names(mouse_DEG_list)
    mouse_DEG_list_transID <- select(org.Mm.eg.db,keys = mouse_DEG_list_name,columns = c("SYMBOL","GENENAME","ENTREZID"),keytype = "ENSEMBL")
    mouse_DEG_list_transID2 <- mapIds(org.Mm.eg.db,keys = mouse_DEG_list_name,column = c("SYMBOL"),keytype = "ENSEMBL")
    ##GO
    ego <- enrichGO(
      gene = row.names(mouse_DEG_list),
      OrgDb = org.Mm.eg.db,
      keyType = "ENSEMBL",
      ont = "CC")
    ego_res <- as.data.frame(ego)
    library(ggplot2)
    library(dplyr)
    ##气泡图
    dotplot(ego,font.size=8)
    ##网络图
    enrichMap(ego, vertex.label.cex=1.2, layout=igraph::layout.kamada.kawai)
    ##GO图
    plotGOgraph(ego)
    ##KEGG
    kegg_list <- mouse_DEG_list_transID[,4]
    kegg <- enrichKEGG(kegg_list,keyType = "kegg",organism = "mmu",pvalueCutoff = 0.05,pAdjustMethod = "BH",qvalueCutoff = 0.1)
    
    dotplot(kegg,font.size=8)
    
    browseKEGG(kegg,'mmu01100')
    
    image.png image.png
    image.png

    参考文章

    1.https://www.jianshu.com/p/fb15249200d7
    2.https://www.jianshu.com/p/fb15249200d7
    3.https://www.jianshu.com/p/ae94178918bc

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