Difference between revisions of "Canavan Task 2 - Sequence alignments"
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**<code> blastpgp -d /mnt/project/pracstrucfunc12/data/big/big_80 -i P45381_wt.fasta -o psiblast_it2_p45381_wt_big80.out -j 2</code> |
**<code> blastpgp -d /mnt/project/pracstrucfunc12/data/big/big_80 -i P45381_wt.fasta -o psiblast_it2_p45381_wt_big80.out -j 2</code> |
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** time: ~2m30 |
** time: ~2m30 |
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− | ** |
+ | ** 500 results |
** best E-Value: E^-142 |
** best E-Value: E^-142 |
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− | ** worst E-Value: |
+ | ** worst E-Value: 3E^-4 |
− | ** |
+ | ** Results with best EValues are mostly Aspartoacylases |
+ | ** New results from 2nd iteration are mostly Succi.. |
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− | ** proteins > EValue e^-15 are often Succinylglutamate Desuccinylases |
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+ | |||
− | ** results similar to blastp run |
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*2 iterations, E-value cutoff 10E-10 |
*2 iterations, E-value cutoff 10E-10 |
Revision as of 18:39, 2 May 2012
Sequence
Native ASPA sequence:
UniProt: P45381
>hsa:443 ASPA, ACY2, ASP; aspartoacylase; K01437 aspartoacylase [EC:3.5.1.15] (A)
MTSCHIAEEHIQKVAIFGGTHGNELTGVFLVKHWLENGAEIQRTGLEVKPFITNPRAVKK
CTRYIDCDLNRIFDLENLGKKMSEDLPYEVRRAQEINHLFGPKDSEDSYDIIFDLHNTTS
NMGCTLILEDSRNNFLIQMFHYIKTSLAPLPCYVYLIEHPSLKYATTRSIAKYPVGIEVG
PQPQGVLRADILDQMRKMIKHALDFIHHFNEGKEFPPCAIEVYKIIEKVDYPRDENGEIA
AIIHPNLQDQDWKPLHPGDPMFLTLDGKTIPLGGDCTVYPVFVNEAAYYEKKEAFAKTTK
LTLNAKSIRCCLH
BLASTP
Run BlastP on student machines with big_80 database.
Command:
blastall -p blastp -d /mnt/project/pracstrucfunc12/data/big/big_80 -i P45381_wt.fasta -o blastp_p45381_wt_big80.out
-p specifies the blast type
-d database
-i input file
-o output file
We got 195 resulting sequences, out of which the last 101 have EValues below E^-15. Most of the resulting proteins are Aspartoacylases of other species. Most of the results with EValue > E^-15 are Succinylglutamate Desuccinylases, which catalyze a reaction similar to Aspartoacylase.
PSIBLAST
Run PsiBlast on student machines with big_80 database.
- 2 iterations, default Evalue 0.002
blastpgp -d /mnt/project/pracstrucfunc12/data/big/big_80 -i P45381_wt.fasta -o psiblast_it2_p45381_wt_big80.out -j 2
- time: ~2m30
- 500 results
- best E-Value: E^-142
- worst E-Value: 3E^-4
- Results with best EValues are mostly Aspartoacylases
- New results from 2nd iteration are mostly Succi..
- 2 iterations, E-value cutoff 10E-10
blastpgp -d /mnt/project/pracstrucfunc12/data/big/big_80 -i P45381_wt.fasta -o psiblast_it2_h10e10_p45381_wt_big80.out -j 2 -h 10e-10
- time ~2m30
- 93 results
- best E Value: E^-145
- worst E Value: 2E-29
- results mainly Aspartoacylases
- 5 iterations, default Evalue 0.002
blastpgp -d /mnt/project/pracstrucfunc12/data/big/big_80 -i P45381_wt.fasta -o psiblast_it5_p45381_wt_big80.out -j 2
- 10 iterations, default Evalue 0.002
blastpgp -d /mnt/project/pracstrucfunc12/data/big/big_80 -i P45381_wt.fasta -o psiblast_it10_p45381_wt_big80.out -j 10
- time: ~10m
- converged after 8 rounds
- 500 results
- best E-Value 5E^-70
- worst E-Value 8E^-38
- most significant results include more Succinylglutamate Desuccinylases than Aspartoacylases
- 10 iterations, E-value cutoff 10E-10
blastpgp -d /mnt/project/pracstrucfunc12/data/big/big_80 -i P45381_wt.fasta -o psiblast_it10_h10e10_p45381_wt_big80.out -j 10 -h 10e-10
HHBLITS
Run HHBlits on student machines with Uniprot20 database.
hhblits -i P45381_wt.fasta -d /mnt/project/pracstrucfunc12/data/hhblits/uniprot20_current -e 0.003 -o hhblits_default.out -E 0.003 -Ofas hhblits_msa_default.txt -n 8