IMGT/V-QUEST programme versions
IMGT/V-QUEST reference directory releases
IMGT/V-QUEST
(V-QUEry and STandardization) [1, 10] at Montpellier is an integrated alignment
tool for the immunoglobulin (IG) and T cell receptor (TR) nucleotide
sequences.
IMGT/V-QUEST compares your germline or rearranged IG or TR variable
sequences with the IMGT/V-QUEST reference directory sets.
The IMGT/V-QUEST ouput displays are according to the IMGT Scientific chart rules and IMGT Repertoire.
Note that IMGT/V-QUEST :
- works with rearranged V-J and V-D-J genes and germline V-GENE, but does not work with germline D-GENE or J-GENE.
- can optionally analyse sequences with DNA insertions or deletions (which do not respect the
IMGT unique numbering). For more information, see Search for insertions and deletions.
- Sequences containing two V domains (as scFv) need to be split (at the level of the linker) prior to IMGT/V-QUEST analysis.
- does not work with out-of-frame pseudogenes as their translated sequences cannot be numbered according to the amino acid
IMGT unique numbering.
You may use BLAST (http://www.imgt.org/blast/) in order
to compare your sequence with F+ORF+all P genes and alleles IMGT reference
sequences (select for « Database » : "IMGT/GENE-DB reference sequences").
- IMGT/V-QUEST does not work, or will give aberrant results, for
too short partial sequences,
sequences containing a cluster of V-GENEs,
or sequences with too long 5'UTR or
3'UTR.
For these sequences,
you may use BLAST (http://www.imgt.org/blast/) in order to identify homologous sequences against IMGT/LIGM-DB (select for « Database » : "IMGT/LIGM-DB") .
Sets of sequences to test IMGT/V-QUEST functionalities
| IMGT/V-QUEST selection and input |
IMGT/V-QUEST is available for IG and TR from different species from the IMGT/V-QUEST Home page.
Select the antigen receptor (Immunoglobulin or T cell receptor) and the species in the IMGT/V-QUEST Home page.
The Search page which appears after your selection shows four sections :
- Your selection
- Nucleotide sequences
- Selection for results
- Advanced parameters
| IMGT/V-QUEST reference directory sets |
- Depending on your selection in the IMGT/V-QUEST Home
page (Immunoglobulin or T cell Receptor, species), your sequence will be
compared to a given IMGT/V-QUEST reference directory set. To see the content
of the available IMGT/V-QUEST reference directory sets, click here.
- The IMGT/V-QUEST
reference directory sets are constituted by sets of sequences which
contain the V-REGION,
D-REGION
and J-REGION
alleles,
isolated from the Functional
and ORF allele IMGT
reference sequences.
-
Since the version Version 2.3.0 of the 5th of July 2007 all "in-frame" Pseudogene
(indicated with (P) following the allele name) and the orphons have been included.
- By definition, these sets contain one sequence
for each allele. Allele names of these sequences are shown in red in Alignments
of alleles.
- Since the Version 3.2.19 of the 20th of June 2011,
gene and allele names are preceded by the short name of the species (encoded on 6 characters, for example Homsap for Homo sapiens
or Musmus for Mus musculus) and followed by the gene and allele
functionality (F for functional, ORF for Open Reading Frame, or P for pseudogene).
Note that parentheses (for example (F)) or brackets (for example [P])
may be associated with
the functionality according to the
IMGT annotation rules for Functionality.
A. Detailed view
Top of the IMGT/V-QUEST result page
The top of the IMGT/V-QUEST result page indicates the number of analysed sequences in the current run.
The name of each sequence is directly linked to its corresponding results.
For each analysed sequence, the IMGT reference directory set used for the alignments
(for example, human IG set) is indicated. The sequence is displayed in FASTA format.
If an input sequence was provided in the antisense orientation,
IMGT/V-QUEST complementary reverses it automatically, and the results will be shown on the complementary reverse sequence
(that is in "sense" orientation for the V-GENE)
.
Result summary
A summary of the results is provided as a table. The Result summary table provides:
-
The functionality
of the sequence, evaluated by IMGT/V-QUEST, is indicated on the first row.
For example: "Productive IGH rearranged sequence (no stop codon and in-frame junction)".
- Note that the functionality of rearranged sequences cannot be determined if the J gene and allele has not been identified. In such cases, the message "No rearrangement found" is indicated.
-
A warning appears in the result summary top line when:
- the V identity percentage is less than 85% and/or when the closest germline
and the analysed sequence show different CDR1-IMGT and/or CDR2-IMGT amino acid lengths: this may indicate potential nucleotide insertion(s) and/or deletion(s)
(see
Somatic hypermutations by insertion or deletion in rearranged cDNA: Human IGHV and [2])
Be aware that a single insertion or deletion will be undetected if the identity percentage is more than 85% and if the CDR1-IMGT length and the CDR2-IMGT length are not altered.
However, they will be visible in 1. Alignment for V-GENE and allele identification, 6. V-REGION alignment,
and 7. V-REGION translation
- the V score is very low (less than 200)
-
The names of the V-GENE and allele and J-GENE and allele are indicated with their alignment score and the percentage of identity.
-
If less than 6 nucleotides of the user sequence are aligned the J-GENE: the J-GENE and allele name is not provided and the message
"Less than 6 nucleotides are aligned" is indicated.
-
A warning may appear with the J-GENE and allele name: this indicates that other possibilities exist for the choice of the J-GENE and allele name.
-
The D-GENE and allele name, the CDR-IMGT lengths and the AA JUNCTION are provided according to the IMGT/JunctionAnalysis results.
-
A warning may appear if IMGT/JunctionAnalysis gives no results. It appears in the "D-GENE and allele"
and "AA JUNCTION" cells.
Note that the letters between parenthesis appear in the order that the warnings are detected.
Results
The results may include (if selected in the input page):
- Alignment for V-GENE and allele identification
- Alignment for D-GENE and allele identification
- Alignment for J-GENE and allele identification
- Results of IMGT/JunctionAnalysis
- Sequence of the JUNCTION ('nt' and 'AA')
- V-REGION alignment
- V-REGION translation
- V-REGION protein display
- V-REGION mutation and AA change table
- V-REGION mutation and AA change statistics
- V-REGION mutation hot spots (motifs and positions in germline V-REGION)
- Sequences of V-, V-J- or V-D-J- REGION ('nt' and 'AA') with gaps in FASTA.
- Annotations by IMGT/Automat
- IMGT Collier de Perles
Example : the sequence of AF184762 accession number.
- Alignments for the V-GENE and allele identification
This
alignment shows your sequence aligned with the closest
V-REGION
alleles from the
IMGT/V-QUEST
reference directory sets.
Dashes indicate identical nucleotides. Dots indicate gaps
according to the
IMGT unique numbering or nucleotides that are not taken into account for the alignments.
- The gene and allele functionality is only shown for pseudogene (P) and "ORF" (ORF) between parentheses.
-
The score is calculated by counting +5 for each nucleotide match and -4
for each mismatch. The score allows to emphasize differences between
the IMGT reference directory sequences and the input sequence, since a
single nucleotide mismatch corresponds to a difference of 9 in the
score.
- For each alignment, the score and the percentage of nucleotide identity are indicated.
The number of identical nucleotides and the total number of nucleotides (excluding gaps)
used for this evaluation are indicated between parentheses.
- Alignments for the D-GENE and allele identification
If this option has been selected in the query page, this
alignment shows your sequence aligned with the closest
D-REGION
alleles from the
IMGT/V-QUEST
reference directory sets.
Dashes indicate identical nucleotides. Dots indicate gaps
according to the
IMGT unique numbering or nucleotides that are not taken into account for the alignments.
-
The score is calculated by counting +5 for each nucleotide match and -4
for each mismatch. The score allows to emphasize differences between
the IMGT reference directory sequences and the input sequence, since a
single nucleotide mismatch corresponds to a difference of 9 in the
score.
- For each alignment, the score and the percentage of nucleotide identity are indicated.
The number of identical nucleotides and the total number of nucleotides (excluding gaps)
used for this evaluation are indicated between parentheses.
- The alignments for the D-REGION and the J-REGION start from the end of
the V-REGION.
-
The "Alignment for D-GENE and allele
identification" provided by IMGT/V-QUEST may show discrepancies with the results obtained
by IMGT/JunctionAnalysis as the way to identify the D-REGIONs is different between the two tools [1,3,4,5]. In case
of discrepancies, the results of IMGT/JunctionAnalysis are the most accurate [4,5].
However, the alignment provided by IMGT/V-QUEST may be useful in some cases for extensive comparison.
- Alignments for the J-GENE and allele identification
.
This
alignment shows your sequence aligned with the closest
J-REGION
alleles from the
IMGT/V-QUEST
reference directory sets.
Dashes indicate identical nucleotides. Dots indicate gaps
according to the
IMGT unique numbering or nucleotides that are not taken into account for the alignments.
-
The score is calculated by counting +5 for each nucleotide match and -4
for each mismatch. The score allows to emphasize differences between
the IMGT reference directory sequences and the input sequence, since a
single nucleotide mismatch corresponds to a difference of 9 in the
score.
- For each alignment, the score and the percentage of nucleotide identity are indicated.
The number of identical nucleotides and the total number of nucleotides (excluding gaps)
used for this evaluation are indicated between parentheses.
- Results of IMGT/JunctionAnalysis
.
Depending on the selection you made in the IMGT/V-QUEST input page, and depending on your sequence, the results of
IMGT/JunctionAnalysis [4] or the
translation of the JUNCTION
will be displayed.
-
A JUNCTION
will extend from 2nd-CYS
104 to J-PHE or
J-TRP
included. J-PHE or
J-TRP
are easily identified when the conserved Phe/Trp-Gly-X-Gly motif of the
J-REGION is present. The translation is displayed up to the motif
(not included) if the motif is present in your sequence or in the closest J-REGION,
or up to the end of your sequence if the motif is not found.
- Analysis of the JUNCTION and Translation of the JUNCTION
If the option "Includes IMGT/JunctionAnalysis" is selected, the results
from IMGT/JunctionAnalysis are displayed.
-
By default, only one D-GENE is searched in the IGH junctions, two in the TRB junctions and three in the TRD junctions.
You are allowed to modify this value in input options (Nb of D-GENE in IGH (or TRB or TRD) JUNCTIONs) of the IMGT/V-QUEST query page.
- The results of IMGT/JunctionAnalysis are detailed in
IMGT/JunctionAnalysis documentation.
-
The number of accepted mutations in 3'V-REGION, D-REGION, and 5'J-REGION corresponds to the default values indicated in
IMGT/JunctionAnalysis documentation except for unmutated IG V-GENE (no mutations in FR1-IMGT, CDR1-IMGT, FR2-IMGT, CDR2-IMGT and FR3-IMGT). In this case, the number of accepted mutations is:
- 0 in 3'V-REGION and 5'J-REGION, and 2 in D-REGION of IGH sequences
- 2 in 3'V-REGION and 5'J-REGION of IGK and IGL sequences
The IMGT/JunctionAnalysis results comprise:
JUNCTION amino acid are colored according to the
IMGT amino acid classes for chemical characteristics [8].
- Eligible D-GENEs
This option "with full list of eligible D-GENEs" is not selected by default in the IMGT/V-QUEST query page.
If selected, IMGT/JunctionAnalysis displays all eligible D-GENEs and alleles
with the length of the germline D-REGION, the sequence identified in the JUNCTION (hyphens indicate similarity), the alignment score and the number of mutations. The column on the right side
indicates the location of the aligned nucleotides in the D-REGION (for example d[3-8]), and the location
of the aligned nucleotides in the JUNCTION (for example s[22-27]);
Note that IMGT/JunctionAnalysis gives no results when:
-
The D gene and allele reference directory of the IGH, TRB or TRD analysed sequences is not managed in IMGT/GENE-DB .
-
The sequence of the 3'V-REGION of the V gene and allele or/and of the 5'J-REGION of the J gene and allele are not well delimitated
(for example in case of partial reference sequences and of reference sequences from cDNA).
-
The number of mutations in the 3'V-REGION, D-REGION, and /or J-REGION is higher than the number set in the "Parameters for IMGT/JunctionAnalysis".
If, in the current conditions,
IMGT/JunctionAnalysis is unable to provide a result, the sequence of the JUNCTION is shown (see below). as well as the JUNCTION nucleotide sequence
formatted for the IMGT/JunctionAnalysis tool.
- Sequence of the JUNCTION ('nt' and 'AA')
The sequence of the JUNCTION is diplayed in nucleotide and in amino acid with the IMGT unique numbering.
- V-REGION alignment
The sequences are
shown with the IMGT
unique numbering and with the IMGT framework region ( FR-IMGT)
and complementarity determining region ( CDR-IMGT)
delimitations. Dashes indicate identical nucleotides. Dots indicate gaps
according to the IMGT unique numbering.

The resulting alignment shows the CDR3-IMGT
of the germline V-REGION
alleles of the IMGT reference directory sets. The CDR3-IMGT
of the input rearranged sequence has to be identified in the
translation of the JUNCTION
(see above).
The correct amino acid numbering of the rearranged CDR3-IMGT is the one shown in the
Results of IMGT/JunctionAnalysis or translation of the JUNCTION
- V-REGION translation
.
The "Translation of the input sequence" shows the nucleotide sequence and
deduced amino acid translation of the input sequence, aligned with the V-REGION of the closest germline V-GENE,
and with the FR-IMGT
and CDR-IMGT
delimitations.
The 3' limit of the CDR3-IMGT
of the input rearranged sequence is correctly identified if the conserved
Phe/Trp-Gly-X-Gly motif of the J-REGION
has been identified. If not, the 3' limit of the CDR3-IMGT
needs to be checked
The correct amino acid numbering of the rearranged CDR3-IMGT is the one shown
in the Results of IMGT/JunctionAnalysis or translation of the JUNCTION

- V-REGION protein display
.
The "V-REGION protein display" shows the
deduced amino acid translation of the input sequence, aligned with the V-REGION of the closest germline V-GENE,
and with the FR-IMGT
and CDR-IMGT
delimitations.
On the third line of the alignment are shown in bold, the input sequence amino acids which different from the closest germline.

- V-REGION mutation and AA change table
.
The "V-REGION mutation and AA change table" shows the mutations of the input sequence by comparison with the V-REGION of the
closest germline V-GENE. Mutations are described according to the IMGT Scientific chart rules and according to
the IMGT unique numbering for
V-REGION. Mutations are shown for FR1-IMGT, CDR1-IMGT, FR2-IMGT, CDR2-IMGT, FR3-IMGT and for the part of the CDR3-IMGT
which can be compared with the V-REGION .
|
IMGT mutation and AA change description
"g36>a" means that the nucleotide "g" at position 36 of the V-REGION of the closest germline V-GENE has been mutated to "a".
If the mutation is nonsilent, the corresponding amino acid change is indicated.
"t88>c, F30>L" means that the nucleotide "t" at position 88 of the V-REGION of the closest germline V-GENE has been
mutated to "c" and that the amino acid "F" at position 30 has been changed to "L".
For each amino acid change, the
IMGT amino acid classes [8] that are conserved despite the amino acid change are
indicated with "+" between parentheses in the following order: hydropathy, volume, chemical characteristics. For example,
F30>L (+ - -) indicates that the two amino acids, F and L, belongs to the same hydropathy class but that the volume and the
chemical characteristics classes are different.
The AA changes are qualified as:
- Very similar: (+ + +)
- Similar: (+ + -) or (+ - +)
- Dissimilar: (+ - -) or (- + -) or (- - +)
- Very dissimilar: (- - -)
As a consequence, two compared AA are qualified as:
- Identical
- Very similar
- Similar
- Dissimilar
- Very dissimilar
|

- V-REGION mutation and AA change statistics
The "V-REGION mutation and AA change statistics" shows the number of mutations and amino acid changes, by comparison with the V-REGION
of the closest germline V-GENE. Numbers of mutations and amino acid changes are shown for the whole V-REGION,
the FR1-IMGT, CDR1-IMGT, FR2-IMGT, CDR2-IMGT, FR3-IMGT and the part of the CDR3-IMGT which can be compared with
the V-REGION.
Note that the number of mutations for the V-REGION and the CDR3-IMGT are calculated with the 3' end delimitation determined
by IMGT/JunctionAnalysis. Numbers added between parentheses
for the V-REGION and the CDR3-IMGT indicate the number of nucleotide differences without taking into account the
shortening of the V-REGION. These numbers include nucleotide differences that result from the N-diversity.
Nucleotides : the number of silent and nonsilent mutations is evaluated, as well as each type of transition and transversion.
Amino acids : The number of identical AA and AA changes is evaluated as well as each type of AA changes.

- V-REGION mutation hot spots
.
The motifs, the positions and the FR-IMGT or CDR-IMGT localisation of hot spots within the closest germline V-REGION are listed.

- Sequences of V-, V-J- or V-D-J- REGION ('nt' and 'AA') with gaps in FASTA.
.
The
sequences of V-REGION and V-D-J or V-J -REGION are displayed with gaps according to the IMGT unique numbering, in nucleotide and in amino acid, in FASTA format.
Amino acid sequences are also displayed on one line.

Note that the V-J- or V-D-J- REGION amino acid
sequences are shown in red for unproductive sequences with an out-of-frame junction.
- Annotations by IMGT/Automat
.
The annotation of the V-REGION, V-J-REGION and /or V-D-J-REGION of the input sequence is provided by IMGT/Automat [9].

- IMGT Collier de Perles for the input sequence V-DOMAIN
.
The
IMGT Collier de Perles
2D graphical representation is automatically generated by the
IMGT/Collier-de-Perles tool program (originally developed by Gérard Mennessier (LPM,
Montpellier, France), Manuel Ruiz, Quentin Kaas and Fran�ois Ehrenmann (LIGM, Montpellier, France)),
The IMGT
IMGT/Collier de Perles for V-DOMAINs are according to the
IMGT unique numbering for V-DOMAIN [7].
IMGT amino acid classes (hydropathy, volume, chemical characteristics) are according to Pommié et al
[8].
Note that: if the option "IMGT Collier de Perles (for a nb of sequences < 5)"
has been selected in "Selection for results display" of the Search page, the Collier de Perles in PNG format is shown (except for TR aplha sequences). Because of several submitted sequences, some IMGT Collier de Perles may wrongly displayed.
IMGT Collier de Perles

B. Synthesis view
Top of the IMGT/V-QUEST result page
The number of analysed sequences is indicated.
Summary Table
The result summary is provided as a table with one row for each input sequence, including:
-
The name of the sequence.
-
The name of the closest V-GENE and allele. Note that a warning may appear with the V-GENE and allele name:
(1) indicates that the V score is very low (less than 200), and/or the V identity percentage is less than 85% and/or when the closest germline and the analysed sequence show different CDR1-IMGT and/or CDR2-IMGT amino acid lengths
The alignment for this sequence has to be checked in A. Detailed view for detection of potential nucleotide insertions and/or deletions,.
-
The functionality
of the sequence. When found, the presence of stop codons is indicated.
-
The V score
-
The percentage of identity
-
The name of the closest J-GENE and allele. Note that two warnings may appear with the J-GENE and allele name:
(a) indicates that other possibilities exist for the choice of the J-GENE and allele name.
-
The D-GENE and allele name, the D reading frame , the CDR-IMGT lengths, the AA JUNCTION and the JUNCTION frame are provided according to the IMGT/JunctionAnalysis results.
In the absence of results of IMGT/JunctionAnalysis, only the AA JUNCTION defined by IMGT/V-QUEST is displayed. Note that a warning may appear to indicate that IMGT/JunctionAnalysis
gives no results. It appears in the "D-GENE and allele"
and "AA JUNCTION" columns for the corresponding sequences.
Results of IMGT/JunctionAnalysis
Links to the results of IMGT/JunctionAnalysis for the junctions are provided .
Alignment with the closest alleles
The names of the closest V-GENE alleles with which your sequences have been aligned are shown, with
the number of sequences aligned to each allele indicated between parentheses.
The name of each allele is directly linked to its corresponding results.
Results for each V-GENE and allele
The results may include (if selected in the input page):
- Alignment for V-GENE
- V-REGION alignment
- V-REGION translation
- V-REGION protein display
- V-REGION protein display with colored AA according to IMGT AA classes
- V-REGION protein display (mutations displayed)
- V-REGION most frequently occurring AA per position and per FR-IMGT and CDR-IMGT
- Results of IMGT/JunctionAnalysis
Example : the sequence of AF184762 accession number.
- Alignment for V-GENE
This
alignment shows your sequences which express the same V aligned with the closest
V-REGION
allele from the
IMGT/V-QUEST
reference directory sets.
Dashes indicate identical nucleotides. Dots indicate gaps
according to the
IMGT unique numbering or nucleotides that are not taken into account for the alignments.
-
Hot spot positions are underlined in the V-REGION or the closest allele.
- The score is indicated for each user sequences.
- The name of the closest J-GENE allele, if found,
is indicated at the end of th alignment
- V-REGION alignment
.
The sequences are
shown with the IMGT
unique numbering and with the IMGT framework region ( FR-IMGT)
and complementarity determining region ( CDR-IMGT)
delimitations. Dashes indicate identical nucleotides. Dots indicate gaps
according to the IMGT unique numbering.
-
Hot spot positions are underlined in the V-REGION or the closest allele.
- The score is indicated for each user sequences.
- The name of the closest J-GENE allele, if found,
is indicated at the end of th alignment

The resulting alignment shows the CDR3-IMGT
of the germline V-REGION
alleles of the IMGT reference directory sets.
- V-REGION translation
.
The "V-REGION Translation " shows the nucleotide sequence and
deduced amino acid translation of the closest V-REGION aligned with the user sequences,
and with the FR-IMGT
and CDR-IMGT
delimitations.
-
Hot spot positions are underlined in the V-REGION or the closest allele.
- The score is indicated for each user sequences.
- The name of the closest J-GENE allele, if found,
is indicated at the end of th alignment

- V-REGION protein display
The "V-REGION protein display" shows the V-REGION protein display of user sequences aligned with the closest V-REGION allele.

- V-REGION protein display (with AA class colors)
.
The "V-REGION protein display" shows the V-REGION protein display of user sequences aligned with the closest V-REGION allele with colored AA according to the AA IMGT classes.

- V-REGION protein display (only AA changes displayed)
The "V-REGION protein display" shows the V-REGION protein display of user sequences aligned with the closest V-REGION allele with only the mutations displayed.

- V-REGION most frequently occurring AA per position and per FR-IMGT and CDR-IMGT
The "V-REGION most frequently occurring AA per position and per FR-IMGT and CDR-IMGT" shows a table, for each FR-IMGT and CDR-IMGT, indicating for each amino acid position the most frequently occurring AA.

- Results of IMGT/JunctionAnalysis
The "Results of IMGT/JunctionAnalysis" shows the results of IMGT/JunctionAnalysis of JUNCTION user sequences per chain type, including Analysis of the JUNCTIONs and Translation of the JUNCTIONs
Note that the "Results of IMGT/JunctionAnalysis" for Synthesis view is available for 'HTML' format only (not for 'Text' format).

C. Excel file
The Excel file comprises 11 sheets:
- The Summary sheet includes:
- Sequence number
- Sequence ID
- Functionality: provides the functionality. A message «(see comment)» may be added. It indicates that a comment related to the functionality has been added in the column «Functionality comment»
- V-GENE and allele: provides the closest V-GENE and allele name(s). A message «(see comment)» may be added. It indicates that a comment related to potential insertions and/or deletions has been added in the column «V-REGION potential ins/del».
- V-REGION score
- V-REGION identity %
- V-REGION identity nt
- V-REGION identity % (with ins/del events): this field is shown if the option «Search for insertions and deletions» is selected.
It indicates the percentage of identity and ratio considering each insertion or deletion as one mutational event.
For each insertion or deletion, whatever its length, "1" is subtracted from the number of identical nucleotides
- V-REGION identity nt (with ins/del events): this field is shown if the option «Search for insertions and deletions» is selected.
It indicates the percentage of identity and ratio considering each insertion or deletion as one mutational event.
For each insertion or deletion, whatever its length, "1" is subtracted from the number of identical nucleotides
- J-GENE and allele: provides the closest J-GENE and allele name(s). A message «(see comment)» may be added. It indicates that a comment related to other possibilities for the J identification has been added in the column «J-GENE and allele comment».
- J-REGION score
- J-REGION identity %
- J-REGION identity nt
- D-GENE and allele: provides the closest D-GENE and allele name(s) identified by IMGT/JunctionAnalysis
- D-REGION reading frame
- CDR1-IMGT length
- CDR2-IMGT length
- CDR3-IMGT length
- CDR-IMGT lengths
- FR-IMGT lengths
- AA JUNCTION (with restored frameshift for out-of-frame junctions, indicated with # in the sequence)
- JUNCTION frame
- Orientation: a sign + indicates «sense» orientation for the cDNA. A sign « indicates «antisens orientation» and therefore, that the sequence was complementary reversed for the IMGT/V-QUEST analysis.
- Functionality comment: explains why a sequence has been identified as «unproductive»
- V-REGION potential ins/del: this field is filled if the option «Search for insertions and deletions» is not selected and if insertions or deletions may be suspected.
- J-GENE and allele comment: this field is filled if other possibilities exist for the choice of the J-GENE and allele
- V-REGION insertions: this field is shown if the option «Search for insertions and deletions» is selected. It indicates the localization of the insertion(s)
- V-REGION deletions: this field is shown if the option «Search for insertions and deletions » is selected. It indicates the localization of the deletion(s)
- Sequence: the user sequence
Note that :
-
if the option "Search for insertions and deletions" is selected and
if insertions have been detected, they appear in capital letters in the user sequence. The results provided
in the other sheets of the Excel file correspond to the IMGT/V-QUEST analysis after removal of the insertions.
-
The AA JUNCTION is not provided only if the JUNCTION can't be analysed by IMGT/JunctionAnalysis.
- The IMGT-gapped-nt-sequences sheet includes:
- Sequence number
- Sequence ID
- Functionality
- V-GENE and allele
- J-GENE and allele
- D-GENE and allele
- Then nt-sequences of:
- V-D-J-REGION : gapped according to the IMGT numbering, if described
- V-J-REGION : gapped according to the IMGT numbering, if described
- V-REGION : gapped according to the IMGT numbering
- FR1-IMGT: gapped according to the IMGT numbering
- CDR1-IMGT: gapped according to the IMGT numbering
- FR2-IMGT: gapped according to the IMGT numbering
- CDR2-IMGT: gapped according to the IMGT numbering
- FR3-IMGT: gapped according to the IMGT numbering
- CDR3-IMGT
- JUNCTION
- J-REGION
- FR4-IMGT
- The nt-sequences sheet includes:
- Sequence number
- Sequence ID
- Functionality
- V-GENE and allele
- J-GENE and allele
- D-GENE and allele
- Then nt-sequences of all constitutive labels of the V-J or V-D-J-REGION
- V-D-J-REGION: if described
- V-J-REGION if described
- V-REGION
- FR1-IMGT
- CDR1-IMGT
- FR2-IMGT
- CDR2-IMGT
- FR3-IMGT
- CDR3-IMGT
- JUNCTION
- labels of the JUNCTION (see 6, Junction sheet section, 'nt sequence of') and (N-D)-REGION
- D-J-REGION
- J-REGION
- FR4-IMGT
- Start and end positions of all constitutive labels of the V-J or V-D-J-REGION
- The IMGT-gapped-AA-sequences sheet includes:
- Sequence number
- Sequence ID
- Functionality
- V-GENE and allele
- J-GENE and allele
- D-GENE and allele
- Then AA-sequences of:
- V-D-J-REGION : gapped according to the IMGT numbering, if described
- V-J-REGION : gapped according to the IMGT numbering, if described
- V-REGION : gapped according to the IMGT numbering
- FR1-IMGT: gapped according to the IMGT numbering
- CDR1-IMGT: gapped according to the IMGT numbering
- FR2-IMGT: gapped according to the IMGT numbering
- CDR2-IMGT: gapped according to the IMGT numbering
- FR3-IMGT: gapped according to the IMGT numbering
- CDR3-IMGT
- JUNCTION
- J-REGION
- FR4-IMGT
- The AA-sequences sheet includes:
- Sequence number
- Sequence ID
- Functionality
- V-GENE and allele
- J-GENE and allele
- D-GENE and allele
- Then, AA-sequences of:
- V-D-J-REGION: if described
- V-J-REGION if described
- V-REGION
- FR1-IMGT
- CDR1-IMGT
- FR2-IMGT
- CDR2-IMGT
- FR3-IMGT
- CDR3-IMGT
- JUNCTION
- J-REGION
- FR4-IMGT
- The Junction sheet includes:
- Sequence number
- Sequence ID
- Functionality
- V-GENE and allele
- J-GENE and allele
- D-GENE and allele
- JUNCTION frame
- Then nt-sequences of labels of the JUNCTION:
- JUNCTION
- JUNCTION with restored frameshift, for out-of-frame junctions
- 3'V-REGION
- P3'V: palindromic (P) nucleotides added downstream (in 3') of the V-REGION
- N-REGION
- N1-REGION
- P5'D: palindromic (P) nucleotides added upstream (in 5') of the D-REGION
- D-REGION
- P3'D: palindromic (P) nucleotides added downstream (in 3') of the D-REGION
- N2-REGION
- P5'J: palindromic (P) nucleotides added upstream (in 5') of the J-REGION
- 5'J-REGION
- Number of nucleotides (nt-nb):
- Sequence number
- JUNCTION nt nb
- 3'V-REGION nt nb
- P3'V nt nb: number of palindromic (P) nucleotides added downstream (in 3') of the V-REGION
- N-REGION nt nb
- N1-REGION nt nb
- P5'D nt nb: number of palindromic (P) nucleotides added upstream (in 5') of the D-REGION
- D-REGION nt nb
- P3'D nt nb: number of palindromic (P) nucleotides added downstream (in 3') of the D-REGION
- N2-REGION nt nb
- P5'J nt nb: number of palindromic (P) nucleotides added downstream (in 5') of the J-REGION
- 5'J-REGION nt nb
- The number of trimmed nt (trimmed-nt nb):
- 3'V-REGION trimmed-nt nb: number of V nucleotides trimmed off the 3' end of the germline V-REGION
- D-REGION 5' trimmed-nt nb: number of D nucleotides trimmed off the 5' end of the germline D-REGION
- D-REGION 3' trimmed-nt nb: number of D nucleotides trimmed off the 3' end of the germline D-REGION
- 5'J-REGION trimmed-nt nb: number of J nucleotides trimmed off the 5' end of germline J-REGION
- The number of mutations (mut-nt nb):
- 3'V-REGION mut-nt nb: number of mutations detected in the 3'V-REGION
- D-REGION mut-nt nb: number of mutations detected in the D-REGION
- 5'J-REGION mut-nt nb: number of mutations detected in the J-REGION
- D-REGION reading frame
- Ngc: gc ratio in nt-sequences encoding N-REGION(s) (N-REGION or N1-REGION+N2-REGION, etc.)
- CDR3-IMGT length
- Molecular mass
- pI
- The parameters used by IMGT/JunctionAnalysis:
- 3'V-REGION accepted-mut nb: number of accepted mutation in the 3'V-REGION, parameter used by IMGT/V-QUEST and IMGT/JunctionAnalysis
- D-REGION accepted-mut nb: number of accepted mutations in the D-REGION, parameter used by IMGT/V-QUEST and IMGT/JunctionAnalysis
- 5'J-REGION accepted mut-nb: number of accepted mutation in the 5'J-REGION, parameter used by IMGT/V-QUEST and IMGT/JunctionAnalysis
- Nb of accepted D-GENE in the JUNCTION for IGH, TRB and TRD loci, parameter used by IMGT/V-QUEST and IMGT/JunctionAnalysis
- CDR3-IMGT : CDR3-IMGT in nucleotides
- CDR3-nt nb : CDR3-IMGT length in nucleotides
- CDR3-IMGT (with frameshift): CDR3-IMGT in nucleotides with restored frameshift (for out-of-frame junctions)
- CDR3-IMGT (AA): CDR3-IMGT in amino acids
- CDR3-IMGT (AA) (with frameshift): CDR3-IMGT in amino acids with restored frameshift (for out-of-frame junctions)
- JUNCTION (AA): JUNCTION in amino acids
- JUNCTION (AA) (with frameshift): JUNCTION in amino acids with restored frameshift (for out-of-frame junctions)
- The V-REGION-mutation-table sheet includes:
- Sequence number
- Sequence ID
- Functionality
- V-GENE and allele
- Then, list of mutations (nt mutation, AA change, AA class identity (+) or change (-), for example t88>c, F30>L(+--)) for:
- V-REGION
- FR1-IMGT
- CDR1-IMGT
- FR2-IMGT
- CDR2-IMGT
- FR3-IMGT
- CDR3-IMGT
- The V-REGION-nt-mutation-statistics sheet includes:
- Sequence number
- Sequence ID
- Functionality
- V-GENE and allele
- Then, for V-REGION, FR1-IMGT, CDR1-IMGT, FR2-IMGT, CDR2-IMGT, FR3-IMGT, CDR3-IMGT:
- Nb of positions including IMGT gaps
- Nb of nucleotides
- Nb of identical nucleotides
- Total nb of mutations
- Nb of silent mutations
- Nb of nonsilent mutations
- The number of transitions
- The number of transversions
- a>c
- c>a
- a>t
- t>a
- g>c
- c>g
- g>t
- t>g
- The V-REGION-AA-mutation-statistics sheet includes:
- Sequence number
- Sequence ID
- Functionality
- V-GENE and allele
- Then, for V-REGION, FR1-IMGT, CDR1-IMGT, FR2-IMGT, CDR2-IMGT, FR3-IMGT, CDR3-IMGT:
- Nb of positions including IMGT gaps (AA)
- Nb of AA
- Nb of identical AA
- Total nb of AA changes
- Nb of AA changes according to AAclassChangeType
- +++
- ++-
- +-+
- +--
- -+-
- --+
- ---
- Nb of AA class changes according to AAclassSimilarityDegree
- Nb of Very similar
- Nb of Similar
- Nb of Dissimilar
- Nb of Very dissimilar
- The V-REGION-mutation-hot-spots sheet includes:
- Sequence number
- Sequence Id
- Functionality
- V-GENE and allele
- Then, Hot spots motifs detected in the closest germline V-REGION with positions and CDR-IMGT and FR-IMGT localization
- (a/t)a
- t(a/t)
- (a/g)g(c/t)(a/t)
- (a/t)(a/g)c(c/t)
- The Parameters sheet includes the date of the analysis, and the general and advanced parameters used by IMGT/V-QUEST:
- Date of the analysis
- IMGT/V-QUEST version
- Species selected for the IMGT reference directory
- Then, selected Advanced parameters:
- IMGT reference directory set
- With allele *01 only : indicated only if that option was selected
- Search for insertions and deletions: yes or no
- Nb of nucleotides to add (or exclude) in 3' of the V-REGION for the evaluation of the alignment score: indicated only if that option was selected
- Nb of nucleotides to exclude in 5' of the V-REGION for the evaluation of the nb of mutations: indicated only if that option was selected.
Results provided following "Search for insertions and deletions" (Advanced parameters)
The option "Search for insertions and/or deletions" of Advanced parameters has to be selected at the bottom of the IMGT/V-QUEST Search page.
Note that:
- A human IG Sequence set to test IMGT/V-QUEST for insertions or deletions is available here
- Insertions or deletions near the 5' end of the V-REGION may not be detected.
Results provided in case of insertions
- in "Detailed view"
-
The detected nucleotide insertions in the submitted sequence by comparison to the IMGT unique numbering
are displayed as capital letters in the FASTA sequence (at the top of results).
- The insertions are described in the Result summary table with:
- their localization in V-REGION
- the display of inserted nucleotides
- the indication if the insertions cause a frameshift
- the V-REGION codon number (according to the IMGT numbering) from which begin the insertions
- the nucleotide position in user submitted sequence from which begin the insertions
- Then IMGT/V-QUEST provides the results of the analysis after removal of the insertions (functionality evaluation,
gene and allele identification, ...).
- In "identity" cell are shown between brackets the percentage of identity and ratio
considering each insertion or deletion as one mutational event.
If (an) insertion(s) is (are) detected, the percentage of identity is calculated after removal of the insertions. Each insertion is
then counted as one mutational event whatever its length. The resulting percentage of identity and ratio are shown between brackets.
- in "Synthesis view"
- IMGT/V-QUEST provides the results of the analysis after removal of the insertions
- A note indicates the sequences for which insertions have been detected. It is strongly recommended to check the description of the insertions in "Detailed view".
- in "Excel file"
Three additional columns are added in the Summary sheet:
- V-REGION identity % (with ins/del events):
it indicates the percentage of identity and ratio considering each insertion or deletion as one mutational event.
For each insertion or deletion, whatever its length, "1" is subtracted from the number of identical nucleotides.
- V-REGION identity nt (with ins/del events):
it indicates the percentage of identity and ratio considering each insertion or deletion as one mutational event.
For each insertion or deletion, whatever its length, "1" is subtracted from the number of identical nucleotides.
- V-REGION insertions: it indicates the localization of the insertion(s).
Note that :
insertions appear in capital letters in the user sequence. The results provided
in the other sheets of the Excel file correspond to the IMGT/V-QUEST analysis after removal of the insertions.
Results provided in case of deletions
- in "Detailed view"
- The deletions are described in the Result summary table with:
- their localization in V-REGION
- the number tof deleted nucleotides
- the indication if the deletions cause a frameshift
- the V-REGION codon number (according to the IMGT numbering) from which begin the deletions
- the nucleotide position in user submitted sequence from which begin the deletions
- Then IMGT/V-QUEST provides the results of the analysis after filling the deletion(s) gap(s) to restore the IMGT numbering (functionality evaluation,
gene and allele idntification, ...)
- In "identity" cell are shown between brackets the percentage of identity and ratio
considering each insertion or deletion as one mutational event.
If (a) deletion(s) is (are) detected, the percentage of identity is calculated after after filling the deletion(s) gap(s). Each deltion is
then counted as one mutational event whatever its length. The resulting percentage of identity and ratio are shown between brackets.
- in "Synthesis view"
- IMGT/V-QUEST provides the results of the analysis after filling the deletion(s) gap(s) to restore the IMGT numbering
- A note indicates the sequences for which deletions have been detected. It is strongly recommended to check the description of the deletions in "Detailed view".
- in "Excel file"
Three additional columns are added in the Summary sheet:
- V-REGION identity % (with ins/del events):
it indicates the percentage of identity and ratio considering each insertion or deletion as one mutational event.
For each insertion or deletion, whatever its length, "1" is subtracted from the number of identical nucleotides.
- V-REGION identity nt (with ins/del events):
it indicates the percentage of identity and ratio considering each insertion or deletion as one mutational event.
For each insertion or deletion, whatever its length, "1" is subtracted from the number of identical nucleotides.
- V-REGION deletions: it indicates the localization of the deletion(s).
References:
| [1] |
Giudicelli, V. et al. Nucl. Acids Res. 32, W435-440 (2004)
PMID: 15215425
LIGM:287 |
| [2] |
Belessi C.J. et al. Eur J Immunol. 36, 1963-74 (2006). |
| [3] |
Giudicelli, V. et al. Stud. Health Technol. Inform. 116, 3-8 (2005) LIGM: 298 |
| [4] |
Yousfi Monod, M. et al. Bioinformatics , 20, i379-i385 (2004)
PMID:
15262823
LIGM:289 |
| [5] |
Lefranc, M.-P. Methods Mol. Biol. 248, 27-49 (2004) PMID: 14970490
LIGM:277
|
| [6] |
Lefranc, M.-P. Current Protocols in Immunology pp. A1W.1-A.1W.15 (2006):311
|
| [7] |
Lefranc, M.-P. et al., Dev. Comp. Immunol., 27, 55-77 (2003)
PMID: 12477501
LIGM:268
|
| [8] |
Pommié, C. et al. J. Mol. Recognit.,
17, 17-32 (2004)
PMID:14872534 LIGM:284 |
| [9] |
Giudicelli, V et al. In: Proceedings of the European Conference on Computational Biology (ECCB 2003), INRIA (DISC/Spid), Paris, DKB-31, pp.103-104.
|
| [10] |
Brochet, X. et al. Nucl. Acids Res. 36, W503-508 (2008).
PMID:18503082 LIGM:344
|
IMGT/V-QUEST Search page
IMGT/V-QUEST Documentation
Contact IMGT/V-QUEST,
Montpellier, France : Véronique Giudicelli (Veronique.Giudicelli@igh.cnrs.fr) and Marie-Paule
Lefranc (Marie-Paule.Lefranc@igh.cnrs.fr)
Created: 07/07/1997
Last updated: 17/09/2012
Authors: Céline Protat, Véronique Giudicelli and Marie-Paule Lefranc.
Marie-Paule.Lefranc@igh.cnrs.fr
Software material and data coming from IMGT server may be used for academic research only,
provided that it is referred to IMGT, and cited as "IMGT, the international ImMunoGeneTics
information system® http://www.imgt.org
(Initiator and coordinator: Marie-Paule Lefranc, Montpellier, France)."
References to cite: Lefranc, M.-P. et al.,
Nucleic Acids Research, 27, 209-212 (1999)
Cover of NAR;
Ruiz, M. et al.,
Nucleic Acids Research, 28, 219-221 (2000);
Lefranc, M.-P.,
Nucleic Acids Research, 29, 207-209 (2001),
Lefranc, M.-P.,
Nucleic Acids Res., 31, 307-310 (2003)
Full text;
Lefranc, M.-P. et al.,
In Silico Biol., 5, 0006 (2004),
Lefranc, M.-P. et al.,
Nucleic Acids Res., 33, D593-D597 (2005)
Full text;
Lefranc, M.-P. et al.,
Nucleic Acids Research 2009 37(Database issue): D1006-D1012; doi:10.1093/nar/gkn838
Full text.
For any other use please contact Marie-Paule Lefranc
Marie-Paule.Lefranc@igh.cnrs.fr.
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