Ensembl BioMart Examples • biomartr
Description: biomartr

## Use Case #1: Functional Annotation of Genes Sharing a Common Evolutionary History

Evolutionary Transcriptomics aims to predict stages or periods of evolutionary conservation in biological processes on the transcriptome level. However, finding genes sharing a common evolutionary history could reveal how the the biological process might have evolved in the first place.

In this `Use Case` we will combine functional and biological annotation obtained with `biomartr` with enriched genes obtained with PlotEnrichment().

### Step 1

For the following example we will use the dataset an enrichment analyses found in PlotEnrichment().

Install and load the myTAI package:

    # install myTAI
    install.packages("myTAI")

    # load myTAI
    library(myTAI)

Download the `Phylostratigraphic Map` of D. rerio:

    # download the Phylostratigraphic Map of Danio rerio
    # from Sestak and Domazet-Loso, 2015

The dataset comes from `Supplementary file 3` of this publication: https://academic.oup.com/mbe/article/32/2/299/1058654#77837069

After downloading `Supplementary file 3`, you will find the file `TableS3-2.xlsx` which can be used in the following `biomartr` functions.

Read the `*.xlsx` file storing the `Phylostratigraphic Map` of D. rerio and format it for the use with `myTAI`:

    # install the readxl package
    install.packages("readxl")

    # load package readxl
    library(readxl)

    # read the excel file
    DrerioPhyloMap.MBEa <- read_excel("TableS3-2.xlsx", sheet = 1, skip = 4)

    # format Phylostratigraphic Map for use with myTAI
    Drerio.PhyloMap <- DrerioPhyloMap.MBEa[ , 1:2]

    # have a look at the final format
    head(Drerio.PhyloMap)

      Phylostrata            ZFIN_ID
    1           1 ZDB-GENE-000208-13
    2           1 ZDB-GENE-000208-17
    3           1 ZDB-GENE-000208-18
    4           1 ZDB-GENE-000208-23
    5           1  ZDB-GENE-000209-3
    6           1  ZDB-GENE-000209-4

Now, `Drerio.PhyloMap` stores the `Phylostratigraphic Map` of D. rerio which is used as background set to perform enrichment analyses with `PlotEnrichment()` from `myTAI`.

### Enrichment Analyses

Now, the `PlotEnrichment()` function visualizes the over- and underrepresented `Phylostrata` of brain specific genes when compared with the total number of genes stored in the `Phylostratigraphic Map` of D. rerio.

    library(readxl)

    # read expression data (organ specific genes) from Sestak and Domazet-Loso, 2015
    Drerio.OrganSpecificExpression <- read_excel("TableS3-2.xlsx", sheet = 2, skip = 3)

    # select only brain specific genes
    Drerio.Brain.Genes <- unlist(unique(na.omit(Drerio.OrganSpecificExpression[ , "brain"])))

    # visualize enriched Phylostrata of genes annotated as brain specific
    PlotEnrichment(Drerio.PhyloMap,
                   test.set     = Drerio.Brain.Genes,
                   measure      = "foldchange",
                   use.only.map = TRUE,
                   legendName   = "PS")

Users will observe that for example brain genes deriving from PS5 are significantly enriched.

Now we can select all brain genes originating in PS5 using the `SelectGeneSet()` function from `myTAI`. Please notice that `SelectGeneSet()` can only be used with phylostratigraphic maps only (`use.map.only = TRUE` argument) since myTAI version > 0.3.0.

    BrainGenes <- SelectGeneSet(ExpressionSet = Drerio.PhyloMap,
                                gene.set      = Drerio.Brain.Genes,
                                use.only.map  = TRUE)

    # select only brain genes originating in PS5
    BrainGenes.PS5 <- BrainGenes[which(BrainGenes[ , "Phylostrata"] == 5), ]

    # look at the results
    head(BrainGenes.PS5)

          Phylostrata           ZFIN_ID
    14851           5 ZDB-GENE-000210-6
    14852           5 ZDB-GENE-000210-7
    14853           5 ZDB-GENE-000328-4
    14856           5 ZDB-GENE-000411-1
    14857           5 ZDB-GENE-000427-4
    14860           5 ZDB-GENE-000526-1

Now users can perform the `biomart()` function to obtain the functional annotation of brain genes originating in PS5.

For this purpose, first we need to find the filter name of the corresponding gene ids such as `ZDB-GENE-000210-6`.

    # find filter for zfin.org ids
    organismFilters("Danio rerio", topic = "zfin_id")

                                name                           description             dataset
    52                  with_zfin_id                       with ZFIN ID(s) drerio_gene_ensembl
    53  with_zfin_id_transcript_name          with ZFIN transcript name(s) drerio_gene_ensembl
    103                      zfin_id ZFIN ID(s) [e.g. ZDB-GENE-060825-136] drerio_gene_ensembl
    274                 with_zfin_id                       with ZFIN ID(s)    drerio_gene_vega
    286                      zfin_id ZFIN ID(s) [e.g. ZDB-GENE-121214-212]    drerio_gene_vega
    366                 with_zfin_id                       with ZFIN ID(s) drerio_gene_ensembl
    367 with_zfin_id_transcript_name          with ZFIN transcript name(s) drerio_gene_ensembl
    417                      zfin_id ZFIN ID(s) [e.g. ZDB-GENE-060825-136] drerio_gene_ensembl
    588                 with_zfin_id                       with ZFIN ID(s)    drerio_gene_vega
    600                      zfin_id ZFIN ID(s) [e.g. ZDB-GENE-121214-212]    drerio_gene_vega
    680                 with_zfin_id                       with ZFIN ID(s) drerio_gene_ensembl
    681 with_zfin_id_transcript_name          with ZFIN transcript name(s) drerio_gene_ensembl
    731                      zfin_id ZFIN ID(s) [e.g. ZDB-GENE-060825-136] drerio_gene_ensembl
    902                 with_zfin_id                       with ZFIN ID(s)    drerio_gene_vega
    914                      zfin_id ZFIN ID(s) [e.g. ZDB-GENE-121214-212]    drerio_gene_vega
                        mart
    52  ENSEMBL_MART_ENSEMBL
    53  ENSEMBL_MART_ENSEMBL
    103 ENSEMBL_MART_ENSEMBL
    274 ENSEMBL_MART_ENSEMBL
    286 ENSEMBL_MART_ENSEMBL
    366 ENSEMBL_MART_ENSEMBL
    367 ENSEMBL_MART_ENSEMBL
    417 ENSEMBL_MART_ENSEMBL
    588 ENSEMBL_MART_ENSEMBL
    600 ENSEMBL_MART_ENSEMBL
    680 ENSEMBL_MART_ENSEMBL
    681 ENSEMBL_MART_ENSEMBL
    731 ENSEMBL_MART_ENSEMBL
    902 ENSEMBL_MART_ENSEMBL
    914 ENSEMBL_MART_ENSEMBL

Now users can retrieve the corresponding GO attribute of D. rerio with `organismAttributes`.

    # find go attribute term for D. rerio
    organismAttributes("Danio rerio", topic = "go")

                                                  name                             description
    33                                           go_id                       GO Term Accession
    36                                 go_linkage_type                   GO Term Evidence Code
    38                            goslim_goa_accession                 GOSlim GOA Accession(s)
    39                          goslim_goa_description                  GOSlim GOA Description
    516                  ggorilla_homolog_ensembl_gene                 Gorilla Ensembl Gene ID
    517  ggorilla_homolog_canomical_transcript_protein      Canonical Protein or Transcript ID
    518               ggorilla_homolog_ensembl_peptide              Gorilla Ensembl Protein ID
    519                    ggorilla_homolog_chromosome                 Gorilla Chromosome Name
    520                   ggorilla_homolog_chrom_start           Gorilla Chromosome Start (bp)
    521                     ggorilla_homolog_chrom_end             Gorilla Chromosome End (bp)
    522                ggorilla_homolog_orthology_type                           Homology Type
    523                       ggorilla_homolog_subtype                                Ancestor
    524          ggorilla_homolog_orthology_confidence    Orthology confidence [0 low, 1 high]
    525                       ggorilla_homolog_perc_id   % Identity with respect to query gene
    526                    ggorilla_homolog_perc_id_r1 % Identity with respect to Gorilla gene
    527                            ggorilla_homolog_dn                                      dN
    528                            ggorilla_homolog_ds                                      dS
    1240                                         go_id                                   GO ID
    1241                                      quick_go                             Quick GO ID
    1370                                         go_id                       GO Term Accession
    1373                               go_linkage_type                   GO Term Evidence Code
    1375                          goslim_goa_accession                 GOSlim GOA Accession(s)
    1376                        goslim_goa_description                  GOSlim GOA Description
    1853                 ggorilla_homolog_ensembl_gene                 Gorilla Ensembl Gene ID
    1854 ggorilla_homolog_canomical_transcript_protein      Canonical Protein or Transcript ID
    1855              ggorilla_homolog_ensembl_peptide              Gorilla Ensembl Protein ID
    1856                   ggorilla_homolog_chromosome                 Gorilla Chromosome Name
    1857                  ggorilla_homolog_chrom_start           Gorilla Chromosome Start (bp)
    1858                    ggorilla_homolog_chrom_end             Gorilla Chromosome End (bp)
    1859               ggorilla_homolog_orthology_type                           Homology Type
    1860                      ggorilla_homolog_subtype                                Ancestor
    1861         ggorilla_homolog_orthology_confidence    Orthology confidence [0 low, 1 high]
    1862                      ggorilla_homolog_perc_id   % Identity with respect to query gene
    1863                   ggorilla_homolog_perc_id_r1 % Identity with respect to Gorilla gene
    1864                           ggorilla_homolog_dn                                      dN
    1865                           ggorilla_homolog_ds                                      dS
    2577                                         go_id                                   GO ID
    2578                                      quick_go                             Quick GO ID
    2707                                         go_id                       GO Term Accession
    2710                               go_linkage_type                   GO Term Evidence Code
    2712                          goslim_goa_accession                 GOSlim GOA Accession(s)
    2713                        goslim_goa_description                  GOSlim GOA Description
    3190                 ggorilla_homolog_ensembl_gene                 Gorilla Ensembl Gene ID
    3191 ggorilla_homolog_canomical_transcript_protein      Canonical Protein or Transcript ID
    3192              ggorilla_homolog_ensembl_peptide              Gorilla Ensembl Protein ID
    3193                   ggorilla_homolog_chromosome                 Gorilla Chromosome Name
    3194                  ggorilla_homolog_chrom_start           Gorilla Chromosome Start (bp)
    3195                    ggorilla_homolog_chrom_end             Gorilla Chromosome End (bp)
    3196               ggorilla_homolog_orthology_type                           Homology Type
    3197                      ggorilla_homolog_subtype                                Ancestor
    3198         ggorilla_homolog_orthology_confidence    Orthology confidence [0 low, 1 high]
    3199                      ggorilla_homolog_perc_id   % Identity with respect to query gene
    3200                   ggorilla_homolog_perc_id_r1 % Identity with respect to Gorilla gene
    3201                           ggorilla_homolog_dn                                      dN
    3202                           ggorilla_homolog_ds                                      dS
    3914                                         go_id                                   GO ID
    3915                                      quick_go                             Quick GO ID
                     dataset                 mart
    33   drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    36   drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    38   drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    39   drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    516  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    517  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    518  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    519  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    520  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    521  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    522  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    523  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    524  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    525  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    526  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    527  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    528  drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1240    drerio_gene_vega ENSEMBL_MART_ENSEMBL
    1241    drerio_gene_vega ENSEMBL_MART_ENSEMBL
    1370 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1373 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1375 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1376 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1853 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1854 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1855 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1856 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1857 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1858 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1859 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1860 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1861 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1862 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1863 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1864 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    1865 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    2577    drerio_gene_vega ENSEMBL_MART_ENSEMBL
    2578    drerio_gene_vega ENSEMBL_MART_ENSEMBL
    2707 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    2710 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    2712 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    2713 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3190 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3191 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3192 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3193 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3194 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3195 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3196 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3197 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3198 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3199 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3200 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3201 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3202 drerio_gene_ensembl ENSEMBL_MART_ENSEMBL
    3914    drerio_gene_vega ENSEMBL_MART_ENSEMBL
    3915    drerio_gene_vega ENSEMBL_MART_ENSEMBL

Now users can specify the filter `zfin_id` and attribute `go_id` to retrieve the GO terms of corresponding gene ids (Please note that this will take some time).

    # retrieve GO terms of D. rerio brain genes originating in PS5
    GO_tbl.BrainGenes <- biomart(genes      = unlist(BrainGenes.PS5[ , "ZFIN_ID"]),
                                 mart       = "ENSEMBL_MART_ENSEMBL",
                                 dataset    = "drerio_gene_ensembl",
                                 attributes = "go_id",
                                 filters    = "zfin_id")

    head(GO_tbl.BrainGenes)

                zfin_id      go_id
    1 ZDB-GENE-000210-6 GO:0060037
    2 ZDB-GENE-000210-6 GO:0046983
    3 ZDB-GENE-000210-7 GO:0046983
    4 ZDB-GENE-000328-4 GO:0007275
    5 ZDB-GENE-000328-4 GO:0007166
    6 ZDB-GENE-000328-4 GO:0035567