Enter four items: query sequence data, taxonomy group information, database file to be searched, and your email address. Then click on "Request for email confirmation".
When your request is successfuly uploaded, a message of "Requested accepted" will appear. Check that your email address is correctly entered.
Request accepted
An email has been sent to xxxxxx@xxxx.xxx.xx for confirmation.
Please click on the link in the email to submit your BlastKOALA job.
Step 2. Job submission
Check your mailbox. You should immediately receive an email from the BlastKOALA server located at the Supercomputer Facility in the Institute for Chemical Research, Kyoto University (scl.kyoto-u.ac.jp).
Your BlastKOALA job request
Query dataset: 3123 entries
KEGG database to be searched: refprok.pep
Please click on the link below to either submit or cancel your job.
https://www.kegg.jp/kegg-bin/blastkoala_submit?id=xx..&type=blastkoala (Submit)
https://www.kegg.jp/kegg-bin/blastkoala_cancel?id=xx..&mode=cancel&type=blastkoala (Cancel)
If no action is taken within 24 hours, your request will be deleted.
Click the submit link in the message to submit your job. Your job will be queued on the BlastKOALA server. The execution time varies depending on how many other jobs are running. Typically, it takes several hours for a search of five thousand sequences against a dataset (such as genus_prokaryotes) of two to three million sequences.
Step 3. Accessing result data
When your job is completed, you will receive a notification email.
Your BlastKOALA job was completed.
Please click on the following link to view the result of annotation.
https://www.kegg.jp/kegg-bin/blastkoala_result?id=xx..&type=blastkoala
Your result data will be removed after seven days.
Click on the link in the message to access your result summary page. You can then view or download the annotation data (K number assingments) and perform KEGG Mapper analysis.
The pie chart shows the functional category defined by the KEGG Orthology system.
Click on "View" in the summary page to open a new page showing your gene list with assigned K numbers, their definitions and the KOALA scores used for the assignments. The second best column contains possible alternatives (K numbers and associated scores), but the interpretation of this column is left to the user.
Query
KO
Definition
HIMB59_00000010
HIMB59_00000020
K00820
glmS; glucosamine--fructose-6-...
HIMB59_00000030
K04042
glmU; bifunctional UDP-N-glucosamine ...
HIMB59_00000040
K07566
tsaC; L-threonylcarbamoyladenylate ...
HIMB59_00000050
The details of K number assignment can be examined for each query gene by clicking on its name in the table. For example, HIMB59_00000020 is linked to the following table containing BLAST hits in the KEGG database searched. This is a simplified version of the GFIT (Gene Function Identification Tool) table internally used by KEGG, here containing only the bit score, identity (%), and overlap (alignment) length, as well as the K number and its definition associated with each similar gene in the database.
Gene (Length)
Score
Ident
Overlap
KO
Definition
bja:bll4607 (608)
466
40
613
K00820
glmS; glucosamine--fructose-6-phosphate...
brs:S23_35090 (608)
459
40
610
K00820
glmS; glucosamine--fructose-6-phosphate...
bju:BJ6T_50800 (608)
455
39
610
K00820
glmS; glucosamine--fructose-6-phosphate...
pre:PCA10_56320 (611)
450
39
616
K00820
glmS; glucosamine--fructose-6-phosphate...
Step 4. Feedback and citation
We welcome your comments and suggestions, especially on any addition or correction of gene/protein function annotation with relevant reference information. Please use the KEGG/GenomeNet Feedback Form.
Please cite the following reference when you make use of BlastKOALA/GhostKOALA.
Kanehisa, M., Sato, Y., and Morishima, K.; BlastKOALA and GhostKOALA: KEGG tools for functional characterization of genome and metagenome sequences. J. Mol. Biol. 428, 726-731 (2016).
[pubmed]
[pdf]
