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Sequence Database Setup: TrEMBL

Trembl Release 39.0
The syntax of the Fasta title line changed (again!) in Trembl release 39.0 (UniProtKB 14.0). This page has been updated to illustrate the new parse rules.

Overview

TrEMBL is a computer-annotated supplement of SWISS-PROT that contains all the translations of EMBL nucleotide sequence entries not yet integrated in SWISS-PROT.

TrEMBL is developed by the SWISS-PROT groups at SIB and EBI.

Download

Expasy: ftp://ftp.expasy.org/databases/uniprot/knowledgebase
EBI: ftp://ftp.ebi.ac.uk/pub/databases/uniprot/knowledgebase

The EBI site mirrors the Expasy site. The relevant files are:

  • Version info: reldate.txt
  • TrEMBL Fasta file: uniprot_trembl.fasta.gz
  • TrEMBL Dat file: uniprot_trembl.dat.gz

To download TrEMBL updates automatically, the relevant definition block in db_update.pl is Trembl_complete_from_EBI.

Taxonomy

Taxonomy for the TrEMBL Dat file is identical to that for SwissProt, and is predefined in mascot.dat as "Swiss-prot DAT".

If you do not wish to have a local copy of the Dat file, and prefer to take taxonomy from the Fasta file instead, you will need to update the taxonomy definition in mascot.dat. Make a backup copy of mascot.dat, then use a text editor to change taxonomy block 3 as follows:

# TAXONOMY FOR SwissProt or Trembl from the fasta file
Taxonomy_3
Identifier SwissProt FASTA
Enabled 1 # 0 to disable it
FromRefFile 0
DescriptionLineSep 0 # ctrl a - hex code '1'. For multiple descriptions per entry
SpeciesFiles NCBI:names.dmp
NodesFiles NCBI:nodes.dmp
DefaultRule NCBI, CHOP:W "OS=\(.*\) PE=" # from release 14.0 onwards
end

Note that mascot.dat must be saved as plain text, so be careful if using a word processor, and ensure the filename is not changed to mascot.dat.txt or something.

Having made these changes, in database maintenance, choose "SwissProt FASTA".

The following taxonomy files are required:

ftp://ftp.ncbi.nih.gov/pub/taxonomy/taxdump.tar.gz
ftp://ftp.ebi.ac.uk/pub/databases/uniprot/knowledgebase/docs/speclist.txt

Note that the taxonomy files go into the taxonomy directory, not into the sequence database directory. Also, some files need to be unpacked (using tar) as well as uncompressed.

Parse Rules

A typical Trembl Fasta title line is:

>tr|A0AQI4|A0AQI4_9ARCH Putative ammonia monooxygenase (Fragment) OS=uncultured archaeon GN=amoA PE=4 SV=1

You can use either the ID (A0AQI4_9ARCH) or the AC (A0AQI4) as the identifier.

ID from Fasta title: ">..|[^|]*|\([^ ]*\)"
AC from Fasta title: ">..|\([^|]*\)"
Description from Fasta title: ">[^ ]* \(.*\)"

The corresponding line in the Dat file is:

ID   A0AQI4_9ARCH              Unreviewed;         206 AA.

ID from Ref file: "^ID   \([^ ]*\)"

Configuration

For this example, the database files were downloaded to C:\Inetpub\MASCOT\sequence\Trembl\current, decompressed using gzip, and renamed to Trembl_39.0.dat and Trembl_39.0.fasta.

When updating an active database, it is important to rename the Fasta file last, because Mascot will begin database exchange as soon as it sees a new Fasta file that matches the wildcard path for the database.

Mascot database maintenance utility

If you decide not to have the reference file locally, full text for individual entries can be retrieved across the web from an SRS server or Expasy. This can be done using either the ID or the AC as the identifier. For Expasy, the syntax for the Path field is:

/cgi-bin/get-sprot-raw.pl?#ACCESSION#

Where #ACCESSION# represents either the AC or ID. For an SRS server, the syntax for the Path field is:

Retrieve by ID: /srsbin/cgi-bin/wgetz?-e+[UNIPROT-id:#ACCESSION#]+-vn+2
Retrieve by AC: /srsbin/cgi-bin/wgetz?-e+[UNIPROT-acc:#ACCESSION#]+-vn+2

This screen shot illustrates a configuration in which the identifier is AC and there is no local Dat file:

Mascot database maintenance utility

If you don't require full text in a Mascot Protein View report, simply leave the Host, Port, and Path fields blank and choose
--- no full text report ---
in the drop down list.

Always test a new definition before applying the changes to mascot.dat.

 
 
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