Bioinformatics for your classroom 1. No programming skills needed 2. Familiarity with personal computer and internet browser NCBI BLAST 3. Customizable and free Seth Bordenstein Discover the Microbes Within! March 12, 2006 Bioinformatics is like using ‘Google’ for DNA sequences Web Access: http://www.ncbi.nlm.nih.gov Web Access Target database: Adjustable using the pull-down menu Primary vs. Derivative Sequence Databases RefSeq Labs Sequencing Centers TATAGCCG AGCTCCGATA CCGATGACAA Curators TATAGCCG TATAGCCG TATAGCCG TATAGCCG Updated continually by NCBI GenBank Updated ONLY by submitters Genome Assembly UniGene Algorithms 50 55 50 40 45 35 40 30 25 Sequence records Total base pairs Release 148: 35 45.2 million records 49.4 billion nucleotides 30 25 20 Average doubling time ≈ 14 months 20 15 15 10 10 5 0 5 ’83 ’84 ’85 ’86 ’87 ’88 ’89 ’90 ’91 ’92 ’93 ’94 ’95 ’96 ’97 ’98 ’99 ’00 ’01 ’02 ’03 ’04 ’05 ’06 0 Total Base Pairs (billions) Sequence Records (millions) 45 LOCUS DEFINITION AY182241 1931 bp mRNA linear PLN 04-MAY-2004 Malus x domestica (E,E)-alpha-farnesene synthase (AFS1) mRNA, complete cds. ACCESSION AY182241 VERSION AY182241.2 GI:32265057 KEYWORDS . SOURCE Malus x domestica (cultivated apple) ORGANISM Malus x domestica Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; Spermatophyta; Magnoliophyta; eudicotyledons; core eudicots; rosids; eurosids I; Rosales; Rosaceae; Maloideae; Malus. REFERENCE 1 (bases 1 to 1931) AUTHORS Pechous,S.W. and Whitaker,B.D. TITLE Cloning and functional expression of an (E,E)-alpha-farnesene synthase cDNA from peel tissue of apple fruit JOURNAL Planta 219, 84-94 (2004) REFERENCE 2 (bases 1 to 1931) AUTHORS Pechous,S.W. and Whitaker,B.D. TITLE Direct Submission JOURNAL Submitted (18-NOV-2002) PSI-Produce Quality and Safety Lab, USDA-ARS, 10300 Baltimore Ave. Bldg. 002, Rm. 205, Beltsville, MD 20705, USA REFERENCE 3 (bases 1 to 1931) AUTHORS Pechous,S.W. and Whitaker,B.D. TITLE Direct Submission JOURNAL Submitted (25-JUN-2003) PSI-Produce Quality and Safety Lab, USDA-ARS, 10300 Baltimore Ave. Bldg. 002, Rm. 205, Beltsville, MD 20705, USA REMARK Sequence update by submitter COMMENT On Jun 26, 2003 this sequence version replaced gi:27804758. FEATURES Location/Qualifiers source 1..1931 /organism="Malus x domestica" /mol_type="mRNA" /cultivar="'Law Rome'" /db_xref="taxon:3750" /tissue_type="peel" gene 1..1931 /gene="AFS1" CDS 54..1784 /gene="AFS1" /note="terpene synthase" /codon_start=1 /product="(E,E)-alpha-farnesene synthase" /protein_id="AAO22848.2" /db_xref="GI:32265058" /translation="MEFRVHLQADNEQKIFQNQMKPEPEASYLINQRRSANYKPNIWK NDFLDQSLISKYDGDEYRKLSEKLIEEVKIYISAETMDLVAKLELIDSVRKLGLANLF EKEIKEALDSIAAIESDNLGTRDDLYGTALHFKILRQHGYKVSQDIFGRFMDEKGTLE DFLHKNEDLLYNISLIVRLNNDLGTSAAEQERGDSPSSIVCYMREVNASEETARKNIK GMIDNAWKKVNGKCFTTNQVPFLSSFMNNATNMARVAHSLYKDGDGFGDQEKGPRTHI LSLLFQPLVN" ORIGIN 1 ttcttgtatc ccaaacatct cgagcttctt gtacaccaaa ttaggtattc actatggaat 61 tcagagttca cttgcaagct gataatgagc agaaaatttt tcaaaaccag atgaaacccg 121 aacctgaagc ctcttacttg attaatcaaa gacggtctgc aaattacaag ccaaatattt 181 ggaagaacga tttcctagat caatctctta tcagcaaata cgatggagat gagtatcgga 241 agctgtctga gaagttaata gaagaagtta agatttatat atctgctgaa acaatggatt // A Traditional GenBank Record Header The Flatfile Format Feature Table Sequence The Header LOCUS DEFINITION ACCESSION VERSION KEYWORDS SOURCE ORGANISM REFERENCE AUTHORS TITLE JOURNAL REFERENCE AUTHORS TITLE JOURNAL REFERENCE AUTHORS TITLE JOURNAL REMARK COMMENT AY182241 1931 bp mRNA linear PLN 04-MAY-2004 Malus x domestica (E,E)-alpha-farnesene synthase (AFS1) mRNA, complete cds. AY182241 AY182241.2 GI:32265057 . Malus x domestica (cultivated apple) Malus x domestica Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; Spermatophyta; Magnoliophyta; eudicotyledons; core eudicots; rosids; eurosids I; Rosales; Rosaceae; Maloideae; Malus. 1 (bases 1 to 1931) Pechous,S.W. and Whitaker,B.D. Cloning and functional expression of an (E,E)-alpha-farnesene synthase cDNA from peel tissue of apple fruit Planta 219, 84-94 (2004) 2 (bases 1 to 1931) Pechous,S.W. and Whitaker,B.D. Direct Submission Submitted (18-NOV-2002) PSI-Produce Quality and Safety Lab, USDA-ARS, 10300 Baltimore Ave. Bldg. 002, Rm. 205, Beltsville, MD 20705, USA 3 (bases 1 to 1931) Pechous,S.W. and Whitaker,B.D. Direct Submission Submitted (25-JUN-2003) PSI-Produce Quality and Safety Lab, USDA-ARS, 10300 Baltimore Ave. Bldg. 002, Rm. 205, Beltsville, MD 20705, USA Sequence update by submitter On Jun 26, 2003 this sequence version replaced gi:27804758. Header: Locus Line LOCUS AY182241 1931 bp mRNA linear PLN 04-MAY-2004 DEFINITION Malus x domestica synthase (AFS1) mRNA, LOCUS AY182241 1931 (E,E)-alpha-farnesene bp mRNA linear PLN 04-MAY-2004 complete cds. ACCESSION AY182241 VERSION AY182241.2 GI:32265057 KEYWORDS . SOURCE Malus x domestica (cultivated apple) ORGANISM Malus x domestica Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; Spermatophyta; Magnoliophyta; eudicotyledons; core eudicots; rosids; eurosids I; Rosales; Rosaceae; Maloideae; Malus. REFERENCE 1 (bases 1 to 1931) AUTHORS Pechous,S.W. and Whitaker,B.D. TITLE Cloning and functional expression of an (E,E)-alpha-farnesene synthase cDNA from peel tissue of apple fruit JOURNAL Planta 219, 84-94 (2004) REFERENCE 2 (bases 1 to 1931) AUTHORS Pechous,S.W. and Whitaker,B.D. TITLE Direct Submission JOURNAL Submitted (18-NOV-2002) PSI-Produce Quality and Safety Lab, USDA-ARS, 10300 Baltimore Ave. Bldg. 002, Rm. 205, Beltsville, MD 20705, USA REFERENCE 3 (bases 1 to 1931) AUTHORS Pechous,S.W. and Whitaker,B.D. TITLE Direct Submission JOURNAL Submitted (25-JUN-2003) PSI-Produce Quality and Safety Lab, USDA-ARS, 10300 Baltimore Ave. Bldg. 002, Rm. 205, Beltsville, MD 20705, USA REMARK Sequence update by submitter COMMENT On Jun 26, 2003 this sequence version replaced gi:27804758. Length Locus name Molecule type Division Modification Date Header: Database Identifiers LOCUS DEFINITION ACCESSION VERSION KEYWORDS SOURCE ORGANISM AY182241 1931 bp mRNA linear PLN 04-MAY-2004 Malus x domestica (E,E)-alpha-farnesene synthase (AFS1) mRNA, Accession complete cds. AY182241 •Stable AY182241.2 GI:32265057 •Reportable . •Universal Malus x domestica (cultivated apple) Malus x domestica Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; Spermatophyta; Magnoliophyta; eudicotyledons; core eudicots; rosids; eurosids I; Rosales; Rosaceae; Maloideae; Malus. 1 (bases 1 to 1931) Pechous,S.W. and Whitaker,B.D. Cloning and functional expression of an (E,E)-alpha-farnesene synthase cDNA from peel tissue of apple fruit Planta 219, 84-94 (2004) 2 (bases 1 to 1931) Pechous,S.W. and Whitaker,B.D. Direct Submission Submitted (18-NOV-2002) PSI-Produce Quality and Safety Lab, USDA-ARS, 10300 Baltimore Ave. Bldg. 002, Rm. 205, Beltsville, MD 20705, USA 3 (bases 1 to 1931) Pechous,S.W. and Whitaker,B.D. Direct Submission Submitted (25-JUN-2003) PSI-Produce Quality and Safety Lab, USDA-ARS, 10300 Baltimore Ave. Bldg. 002, Rm. 205, Beltsville, MD 20705, USA Sequence update by submitter On Jun 26, 2003 this sequence version replaced gi:27804758. ACCESSION AY182241 VERSION AY182241.2 REFERENCE AUTHORS TITLE JOURNAL REFERENCE AUTHORS TITLE JOURNAL REFERENCE AUTHORS TITLE JOURNAL REMARK COMMENT GI:32265057 Header: Organism LOCUS DEFINITION AY182241 1931 bp mRNA linear PLN 04-MAY-2004 Malus x domestica (E,E)-alpha-farnesene synthase (AFS1) mRNA, complete cds. ACCESSION AY182241 VERSION AY182241.2 GI:32265057 KEYWORDS . SOURCE Malus x domestica (cultivated apple) SOURCE (cultivated apple) ORGANISMMalus Malusxx domestica domestica ORGANISM Malus x domestica Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; Spermatophyta; Magnoliophyta; Streptophyta; eudicotyledons; core eudicots; Eukaryota; Viridiplantae; Embryophyta; rosids; eurosids I; Rosales; Rosaceae; Maloideae; Malus. Tracheophyta; Spermatophyta; Magnoliophyta; eudicotyledons; REFERENCE 1 (bases 1 to 1931) eudicots; eurosids I; Rosales; Rosaceae; AUTHORS core Pechous,S.W. androsids; Whitaker,B.D. TITLE Maloideae; Cloning andMalus. functional expression of an (E,E)-alpha-farnesene synthase cDNA from peel tissue of apple fruit JOURNAL Planta 219, 84-94 (2004) REFERENCE 2 (bases 1 to 1931) AUTHORS Pechous,S.W. and Whitaker,B.D.NCBI-controlled taxonomy TITLE Direct Submission JOURNAL Submitted (18-NOV-2002) PSI-Produce Quality and Safety Lab, USDA-ARS, 10300 Baltimore Ave. Bldg. 002, Rm. 205, Beltsville, MD 20705, USA REFERENCE 3 (bases 1 to 1931) AUTHORS Pechous,S.W. and Whitaker,B.D. TITLE Direct Submission JOURNAL Submitted (25-JUN-2003) PSI-Produce Quality and Safety Lab, USDA-ARS, 10300 Baltimore Ave. Bldg. 002, Rm. 205, Beltsville, MD 20705, USA REMARK Sequence update by submitter COMMENT On Jun 26, 2003 this sequence version replaced gi:27804758. The Feature Table FEATURES source gene CDS start (atg) Coding sequence Location/Qualifiers 1..1931 /organism="Malus x domestica" /mol_type="mRNA" /cultivar="'Law Rome'" /db_xref="taxon:3750" /tissue_type="peel" 1..1931 /gene="AFS1" stop (tag) 54..1784 /gene="AFS1" /note="terpene synthase" /codon_start=1 /product="(E,E)-alpha-farnesene synthase" /protein_id="AAO22848.2" /db_xref="GI:32265058" /translation="MEFRVHLQADNEQKIFQNQMKPEPEASYLINQRRSANYKPNIWK NDFLDQSLISKYDGDEYRKLSEKLIEEVKIYISAETMDLVAKLELIDSVRKLGLANLF EKEIKEALDSIAAIESDNLGTRDDLYGTALHFKILRQHGYKVSQDIFGRFMDEKGTLE NHHFAHLKGMLELFEASNLGFEGEDILDEAKASLTLALRDSGHICYPDSNLSRDVVHS LELPSHRRVQWFDVKWQINAYEKDICRVNATLLELAKLNFNVVQAQLQKNLREASRWW ANLGIADNLKFARDRLVECFACAVGVAFEPEHSSFRICLTKVINLVLIIDDVYDIYGS EEELKHFTNAVDRWDSRETEQLPECMKMCFQVLYNTTCEIAREIEEENGWNQVLPQLT KVWADFCKALLVEAEWYNKSHIPTLEEYLRNGCISSSVSVLLVHSFFSITHEGTKEMA DFLHKNEDLLYNISLIVRLNNDLGTSAAEQERGDSPSSIVCYMREVNASEETARKNIK GMIDNAWKKVNGKCFTTNQVPFLSSFMNNATNMARVAHSLYKDGDGFGDQEKGPRTHI The Sequence: What do you do with it? ORIGIN // 1 61 121 181 ttcttgtatc tcagagttca aacctgaagc ggaagaacga ccaaacatct cttgcaagct ctcttacttg tttcctagat cgagcttctt gataatgagc attaatcaaa caatctctta gtacaccaaa agaaaatttt gacggtctgc tcagcaaata ttaggtattc tcaaaaccag aaattacaag cgatggagat actatggaat atgaaacccg ccaaatattt gagtatcgga 1741 1801 1861 1921 ggacccacat aataaatagc tgtaacgttg aaaaaaaaaa cctgtcttta ctattccaac ctcttgtaaa ctagtactca tatagtttga agcaaaagtt tgcggttcag ttcgtcatgg ataaattaat ctttacagtt ttgccaaaga ttatgaataa aaagttgtag tttgtcgttt aaaaaaaaaa a Uses of BLAST: Query a database for sequences similar to an input sequence. Identify previously characterized sequences. Find phylogenetically related sequences. Identify possible functions based on similarities to known sequences. The different versions of BLAST Advantages of using an online exercise for Bioinformatics: • The real experience; Interactive for self-inquiry • Continuously updateable with suggestions • Links education with technology • Uses the computer - a friendly tool to students • Free and no supply costs if you have computers • You can start this tutorial on Monday! Let’s Begin Our Bioinformatic Exercise! http://serc.carleton.edu/microbelife/k12/bioinformatics/module1.html