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NCBI Molecular Biology Resources
Part 2
November 2008 Peter Cooper
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• Genomic Resources
• NCBI BLAST
NCBI Resources: Part 2
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Genome Resources
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Complete Genomes including draft assemblies, Oct 2008
Organisms: Viruses (2,187) Archaea (60) Bacteria (1,284) Eukaryotes (191)Organelles: Mitochondria (1,537)Plastids (147)
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Higher Eukaryotic Genomes Oct 2008
• Animals (78)– Placozoa (1)– Cnidaria (2)– Nematodes (7)– Mollusks (1)– Arthropods (23)
• Insects (21)• Crustaceans (1)• Arachnids (1)
– Echinoderms (1)– Chordates (42)
• Fungi (57)– Ascomycetes (58)– Basidiomycetes (9)
• Land Plants– Angiosperms (7)– Mosses (1)
metazoa[organism] OR dikarya[organism] OR streptophyta[organism]
153 Total species
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Genome Resources: All Genomes
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Eukaryotic Genomes Only
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Microbial Genomes Only: COGs and Protein Clusters
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Selected Eukaryotic Genomes
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NM_000249: Genome Links
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Map Viewer: MLH1CustomizableCustomizable
NCBI Assembly
EST Hits
Gene Annotations
Models
Transcripts
Download data and sequencesDownload data and sequences
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eSynteny: Mammalian Genomes
Albumin Gene FamilyAlbumin Gene Family
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The New Homologene
early globin gene
A-chain gene B-chain gene
frog A chick A mouse A mouse B chick B frog B
paralogsorthologs orthologs
gene duplication• No longer UniGene based• Protein similarities first• Guided by taxonomic tree• Includes orthologs and paralogs
• No longer UniGene based• Protein similarities first• Guided by taxonomic tree• Includes orthologs and paralogs
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Finding Homologs: HomoloGene
GeneGene
Provides Neighboring Function Gene
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HomoloGene Cluster
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Expanded Coverage: UniGene
Fathead Minnow MLH1Fathead Minnow MLH1
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Microbial Genomes
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E. coli mutL Gene Record
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Entrez Genomes View
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eNew Sequence Viewer (All
Genomes)
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Incipient Genome Browser
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COGs AnalysisE.Coli K12 Genome
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Protein Clusters (Update for COGs)
Genomic order
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Sequence Similarity Searching
Basic Local Alignment Search Tool
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The Flavors of BLAST• Position independent scoring
– Standard BLAST• traditional contiguous word hit• nucleotide, protein and translations
– Megablast• can use discontiguous words• nucleotide only• optimized for large batch searches
• Position dependent scoring
– PSI-BLAST• constructs PSSMs automatically• searches protein database with PSSMs
– RPS BLAST• searches a database of PSSMs• basis of conserved domain database
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Basic BLAST: Databases
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BLAST Databases: Non-redundant protein
nr (non-redundant protein sequences)– GenBank CDS translations– NP_, XP_ RefSeqs– Outside Protein
• PIR, Swiss-Prot, PRF• PDB (sequences from structures)
pat protein patents
env_nr environmental samples
nr (non-redundant protein sequences)– GenBank CDS translations– NP_, XP_ RefSeqs– Outside Protein
• PIR, Swiss-Prot, PRF• PDB (sequences from structures)
pat protein patents
env_nr environmental samples
Servicesblastpblastx
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eNucleotide Databases: Human and
Mouse
• Human and mouse genomic and transcript now default• Separate sections in output for mRNA and genomic• Direct links to Map Viewer for genomic sequences
Megablast, blastn service
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Nucleotide Databases: Traditional
Servicesblastntblastntblastx
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Nucleotide Databases: Traditional
• nr (nt)– Traditional GenBank– NM_ and XM_
RefSeqs• refseq_rna
• refseq_genomic– NC_ RefSeqs
• dbest – EST Division
• est_human, mouse, others
• htgs – HTG division
• gss – GSS division
• wgs– whole genome
shotgun
• env_nt– environmental
samples
Databases are mostly non-overlapping
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WWW BLAST
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eThe BLAST homepage
New URL: http://blast.ncbi.nlm.nih.gov/New URL: http://blast.ncbi.nlm.nih.gov/
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Universal Form: Protein
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Universal Form: Nucleotide
SpeedSpeedSensitivitySensitivity
More
Less
Less
More
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Limiting Database: Organism
Organism autocompleteOrganism autocomplete
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Limiting Database: Entrez Query
all[filter] NOT mammals[organism]
gene_in_mitochondrion[Properties]2006:2007 [Modification Date]
Nucleotidebiomol_mrna[Properties]biomol_genomic[Properties]
all[filter] NOT mammals[organism]
gene_in_mitochondrion[Properties]2006:2007 [Modification Date]
Nucleotidebiomol_mrna[Properties]biomol_genomic[Properties]
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eAlgorithm parameters: Protein
Adjust to set stringencyAdjust to set stringency
May limit resultsMay limit results
Default statistics adjustmentfor compositional bias
Default statistics adjustmentfor compositional bias
Off now by default. Conflicts withcomp-based statsOff now by default. Conflicts withcomp-based stats
ExpandExpand
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Automatic Short Sequence Adjustment
e-value 20000Word Size 2Matrix PAM30Comp Stats OffLow Comp Filter Off
Nucleotide and Protein
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Algorithm parameters: Nucleotideblastnblastn
•Masks species-specific interspersed repeats•Essential for genomic query sequences•Masks species-specific interspersed repeats•Essential for genomic query sequences
•Prevents starting alignment in masked region•Allows extensions through masked regions•Prevents starting alignment in masked region•Allows extensions through masked regions
Masks LC sequence (simple repeats)Masks LC sequence (simple repeats)
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BLAST Formatting Options
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Formatting Page (Now on Results)
Alignment ViewPairwisePairwise with dots for identitiesQuery-anchored with dots for identitiesQuery-anchored with letters for identitiesFlat query-anchored with dots for identitiesFlat-query anchored with letters for identities
Alignment ViewPairwisePairwise with dots for identitiesQuery-anchored with dots for identitiesQuery-anchored with letters for identitiesFlat query-anchored with dots for identitiesFlat-query anchored with letters for identities
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eDownload Options (Now on
Results)
Structured Formats
Saved Settings•Reusable on Web•Portable to Standalone
PSSM•Reusable on Web•Portable to Standalone
Standalone formatter (future)
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Structured formats: XML and ASN.1
<Iteration_hits>−<Hit><Hit_num>1</Hit_num><Hit_id>gi|730028|sp|P40692|MLH1_HUMAN</Hit_id>−<Hit_def>DNA mismatch repair protein Mlh1 (MutL protein homolog 1)</Hit_def><Hit_accession>P40692</Hit_accession><Hit_len>756</Hit_len>−<Hit_hsps>−<Hsp><Hsp_num>1</Hsp_num><Hsp_bit-score>1568.9</Hsp_bit-score><Hsp_score>4061</Hsp_score><Hsp_evalue>0</Hsp_evalue><Hsp_query-from>1</Hsp_query-from><Hsp_query-to>756</Hsp_query-to><Hsp_hit-from>1</Hsp_hit-from><Hsp_hit-to>756</Hsp_hit-to><Hsp_query-frame>0</Hsp_query-frame><Hsp_hit-frame>0</Hsp_hit-frame><Hsp_identity>0</Hsp_identity><Hsp_positive>0</Hsp_positive><Hsp_gaps>0</Hsp_gaps><Hsp_align-len>756</Hsp_align-len>
Seq-annot ::= { desc { user { type str "Hist Seqalign" , data { { label str "Hist Seqalign" , data bool TRUE } } } , user { type str "Blast Type" , data { { label id 0 , data int 0 } } } , user { type str "BLAST database title" , data { { label str "Non-redundant SwissProt
Seq-annot ::= { desc { user { type str "Hist Seqalign" , data { { label str "Hist Seqalign" , data bool TRUE } } } , user { type str "Blast Type" , data { { label id 0 , data int 0 } } } , user { type str "BLAST database title" , data { { label str "Non-redundant SwissProt
XMLXML
ASN.1ASN.1
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The Hit Table# BLASTP 2.2.17 (Aug-26-2007)# Query: gi|4557757|ref|NP_000240.1| MutL protein homolog 1 [Homo sapiens]# Database: swissprot# Fields: query id, subject ids, % identity, % positives, alignment length, mismatches, gap opens, q. start, q. end, s. start, s. end, evalue, bit score# 80 hits foundref|NP_000240.1||gi|4557757 gi|1709056|sp|P38920|MLH1_YEAST 36.68 56.91 796 426 18 8 756 5 769 7e-138 491ref|NP_000240.1||gi|4557757 gi|48474996|sp|Q9P7W6|MLH1_SCHPO 37.24 54.04 768 371 16 8 756 9 684 8e-122 437ref|NP_000240.1||gi|4557757 gi|25090753|sp|Q8RA70|MUTL_THETN 37.44 54.62 390 231 7 8 394 4 383 5e-59 229ref|NP_000240.1||gi|4557757 gi|25090732|sp|Q8KAX3|MUTL_CHLTE 35.95 54.05 370 229 5 8 375 4 367 5e-55 215ref|NP_000240.1||gi|4557757 gi|127552|sp|P23367.2|MUTL_ECOLI 35.99 58.11 339 202 7 8 334 3 338 8e-55 214ref|NP_000240.1||gi|4557757 gi|29427778|sp|Q8FAK9|MUTL_ECOL6 35.99 58.11 339 202 7 8 334 3 338 1e-54 214ref|NP_000240.1||gi|4557757 gi|20455084|sp|Q8XDN4|MUTL_ECO57 35.99 58.11 339 202 7 8 334 3 338 1e-54 214ref|NP_000240.1||gi|4557757 gi|59798328|sp|Q72PF7|MUTL_LEPIC 36.27 55.20 375 221 8 6 375 2 363 3e-54 213ref|NP_000240.1||gi|4557757 gi|13431695|sp|P57886|MUTL_PASMU 35.48 58.94 341 213 6 8 345 3 339 4e-54 212ref|NP_000240.1||gi|4557757 gi|1171080|sp|P44494|MUTL_HAEIN 35.74 59.87 319 198 6 8 323 3 317 5e-54 212ref|NP_000240.1||gi|4557757 gi|20455102|sp|Q8ZIW4|MUTL_YERPE 36.01 58.63 336 207 6 8 339 3 334 6e-54 212ref|NP_000240.1||gi|4557757 gi|20455152|sp|Q9JYT2|MUTL_NEIMB 33.96 55.35 374 224 8 8 376 4 359 2e-53 210ref|NP_000240.1||gi|4557757 gi|20139217|sp|Q9KAC1|MUTL_BACHD 35.39 55.90 356 214 6 8 362 4 344 2e-53 209ref|NP_000240.1||gi|4557757 gi|31076794|sp|Q87L05|MUTL_VIBPA 35.33 58.38 334 210 5 8 338 3 333 3e-53 209ref|NP_000240.1||gi|4557757 gi|20455150|sp|Q9JTS2|MUTL_NEIMA 36.94 58.28 314 183 5 8 316 4 307 5e-53 209ref|NP_000240.1||gi|4557757 gi|56749233|sp|Q6GHD9|MUTL_STAAR 38.28 58.46 337 193 7 6 335 2 330 1e-52 207ref|NP_000240.1||gi|4557757 gi|25090739|sp|Q8NWX9|MUTL_STAAW 38.28 58.46 337 193 7 6 335 2 330 1e-52 207ref|NP_000240.1||gi|4557757 gi|71151979|sp|Q5HGD5|MUTL_STAAC 38.28 58.46 337 193 7 6 335 2 330 1e-52 207ref|NP_000240.1||gi|4557757 gi|54037875|sp|P65492|MUTL_STAAN 38.28 58.46 337 193 7 6 335 2 330 2e-52 207ref|NP_000240.1||gi|4557757 gi|20043258|sp|Q9KV13|MUTL_VIBCH 35.74 58.56 333 204 6 8 335 3 330 2e-52 207ref|NP_000240.1||gi|4557757 gi|127553|sp|P14161|MUTL_SALTY 35.10 56.93 339 205 7 8 334 3 338 3e-52 206ref|NP_000240.1||gi|4557757 gi|20455140|sp|Q9CDL1|MUTL_LACLA 36.31 56.55 336 196 5 6 334 2 326 4e-52 206ref|NP_000240.1||gi|4557757 gi|61214242|sp|Q7MH01|MUTL_VIBVY 34.63 58.51 335 213 5 8 339 3 334 4e-52 206ref|NP_000240.1||gi|4557757 gi|20455099|sp|Q8Z187|MUTL_SALTI 35.10 56.93 339 205 7 8 334 3 338 4e-52 206ref|NP_000240.1||gi|4557757 gi|31076809|sp|Q8DCV0|MUTL_VIBVU 34.63 58.51 335 213 5 8 339 3 334 6e-52 205ref|NP_000240.1||gi|4557757 gi|71648717|sp|Q5E2C6|MUTL_VIBF1 36.71 59.81 316 186 6 8 316 3 311 1e-51 204ref|NP_000240.1||gi|4557757 gi|37999611|sp|Q88DD1|MUTL_PSEPK 30.34 48.97 435 278 7 8 419 7 439 2e-51 203
Also available in comma separated format for ExcelAlso available in comma separated format for Excel
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PSSMs: Restart PSI-BLAST
ASCII encoded, Web onlyASCII encoded, Web only
ASN.1 ScoreMat, PortableASN.1 ScoreMat, Portable
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BLAST TreeView
Black bear mt genome vs. RefSeq GenomicBlack bear mt genome vs. RefSeq Genomic
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Distance Tree Carnivore Mitochondrial Genome
bearswalrus fur seal
sea lions
true seals
dogsmongooses
catsred pandaweasels
raccoon
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Genome and Specialized BLAST
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eNucleotide Databases: Human and
Mouse
• Human and mouse genomic and transcript now default• Separate sections in output for mRNA and genomic• Direct links to Map Viewer for genomic sequences
Megablast, blastn service
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Genome BLAST pages
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Map Viewer Homepage
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Poplar Genome BLAST
![Page 53: NCBI FieldGuide NCBI Molecular Biology Resources Part 2 November 2008 Peter Cooper](https://reader036.vdocuments.us/reader036/viewer/2022062409/56649ea95503460f94bae285/html5/thumbnails/53.jpg)
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tblastn Genome BLAST Results
Protein-nucleotide alignmentsProtein-nucleotide alignments
Exons and genes mixedExons and genes mixed
![Page 54: NCBI FieldGuide NCBI Molecular Biology Resources Part 2 November 2008 Peter Cooper](https://reader036.vdocuments.us/reader036/viewer/2022062409/56649ea95503460f94bae285/html5/thumbnails/54.jpg)
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eGenomic Context of BLAST Hits
![Page 55: NCBI FieldGuide NCBI Molecular Biology Resources Part 2 November 2008 Peter Cooper](https://reader036.vdocuments.us/reader036/viewer/2022062409/56649ea95503460f94bae285/html5/thumbnails/55.jpg)
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Hits in Map Viewer
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Specialized BLAST Pages
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BLAST URL APIhttp://www.ncbi.nlm.nih.gov/BLAST/Doc/urlapi.htmlhttp://www.ncbi.nlm.nih.gov/BLAST/Doc/urlapi.html
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eBLAST: standalone, clients,
databases
ftp> open ftp.ncbi.nih.gov..ftp> cd blast
ftp> open ftp.ncbi.nih.gov..ftp> cd blast
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Standalone BLASTC toolkit BLAST C++ toolkit BLAST
BLAST
E:\Blast\bin>blastall –i purf.fsa –d swissprot –p blastp
BLAST+
E:\blast+\bin>blastp –query purf.fsa –db swissprot
BLAST
E:\Blast\bin>blastall –i purf.fsa –d swissprot –p blastp
BLAST+
E:\blast+\bin>blastp –query purf.fsa –db swissprot
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Service Addresses
•General Help [email protected]•BLAST [email protected]
Telephone support: 301- 496- 2475