Presentation is loading. Please wait.

Presentation is loading. Please wait.

Introduktion til Bioinformatik Hold 01 Oktober 2010.

Similar presentations


Presentation on theme: "Introduktion til Bioinformatik Hold 01 Oktober 2010."— Presentation transcript:

1 Introduktion til Bioinformatik Hold 01 Oktober 2010

2 Introduktion Rasmus Wernersson, Lektor Anders Gorm Pedersen, Docent Center for Biologisk Sekvensanalyse, DTU

3 Oversigt Data & Databaser Metoder Taksonomi DNA Protein Protein struktur Alignment Pairwise + Multiple BLAST (søgning) Fylogenetiske træer PyMOL (3D visualisering) Opsamlende øvelse Malaria vaccine

4 Øvelserne er det primære

5 Kursusplan på vores wiki

6 On evolution and sequences Background information

7 Classification: Linnaeus Carl Linnaeus

8 Classification: Linnaeus Hierarchical system –Kingdom –Phylum –Class –Order –Family –Genus –Species

9 Classification depicted as a tree

10 No “mixed” animals Source:

11 Classification depicted as a tree SpeciesGenusFamilyOrderClass

12 Comparison of limbs Image source:

13 Theory of evolution Charles Darwin

14 Phylogenetic basis of systematics Linnaeus: Ordering principle is God. Darwin: Ordering principle is shared descent from common ancestors. Today, systematics is explicitly based on phylogeny.

15 Natural Selection: Darwin’s four postulates More young are produced each generation than can survive to reproduce. Individuals in a population vary in their characteristics. Some differences among individuals are based on genetic differences. Individuals with favorable characteristics have higher rates of survival and reproduction. Evolution by means of natural selection Presence of ”design-like” features in organisms: Quite often features are there “for a reason”

16 Evolution at the sequence level

17 About DNA DNA contains the recipes of how to make protein / enzymes. Every time a cells divides it’s DNA is duplicated, and each daughter cell gets a copy.

18 The DNA alphabet The information in the DNA is written in a four letter code: A, T, G, C. The DNA can be “sequenced” and the result stored in a computer file. ATGGCCCTGTGGAT

19 DNA is always written 5’  3’ 5’ AGCC 3’ 3’ TCGG 5’ 5’ ATGGCCAGGTAA 3’ DNA backbone: (Deoxy)ribose: Ribose Deoxyribose ’ 3’ 5’ 3’

20 ATGGCCCTGTGGATGCG Can DNA be changed?

21 ATGGCCCTGTGGATGCG ATGGCCCTATGGATGCG Can DNA be changed?

22 A history of mutations ATGGCCCTGTGTATGCG ATGGCAATGTGGATGCA ATGGCCCTGTGGATGCG ATGGCCCCGTGGATGCG ATGTCCCCGTGGATGCG ATGGCCCCGTGGAACCG Time

23 Species1: ATGGCAATGTGGATGCA Species2: ATGGCCCCGTGGAACCG Species3: ATGTCCCCGTGGATGCG “DNA alignment” 3 6 5

24 Real life example: Alignment Insulin from 7 different species Homo:ATGGCCCTGTGGATGCGCCTCCTGCCCCTGCTGGCGCTGCTGGCCCTCTGGGGACCTGACCCAGCCGCAGCCTTTGTGAA Pan:ATGGCCCTGTGGATGCGCCTCCTGCCCCTGCTGGTGCTGCTGGCCCTCTGGGGACCTGACCCAGCCTCGGCCTTTGTGAA Sus:ATGGCCCTGTGGACGCGCCTCCTGCCCCTGCTGGCCCTGCTGGCCCTCTGGGCGCCCGCCCCGGCCCAGGCCTTCGTGAA Ovis:ATGGCCCTGTGGACACGCCTGGTGCCCCTGCTGGCCCTGCTGGCACTCTGGGCCCCCGCCCCGGCCCACGCCTTCGTCAA Canis:ATGGCCCTCTGGATGCGCCTCCTGCCCCTGCTGGCCCTGCTGGCCCTCTGGGCGCCCGCGCCCACCCGAGCCTTCGTTAA Mus:ATGGCCCTGTTGGTGCACTTCCTACCCCTGCTGGCCCTGCTTGCCCTCTGGGAGCCCAAACCCACCCAGGCTTTTGTCAA Gallus:ATGGCTCTCTGGATCCGATCACTGCCTCTTCTGGCTCTCCTTGTCTTTTCTGGCCCTGGAACCAGCTATGCAGCTGCCAA

25 Real life example: Tree

26 Interpretation of Multiple Alignments Conserved features assumed to be important for functionality For instance: conserved pairs of cysteines indicate possible disulphide bridge

27 Darwin: all organisms are related through descent with modification Prediction: similar molecules have similar functions in different organisms Protein synthesis carried out by very similar RNA-containing molecular complexes (ribosomes) that are present in all known organisms Sequences are related

28 Sequences are related, II Related oxygen- binding proteins in humans

29 DNA as Biological Information Rasmus Wenersson

30 Overview Learning objectives –About Biological Information –A note about DNA sequencing techniques and DNA data –File formats used for biological data –Introduction to the GenBank database

31 Information flow in biological systems

32 DNA sequences = summary of information 5’ AGCC 3’ 3’ TCGG 5’ 5’ ATGGCCAGGTAA 3’ DNA backbone: (Deoxy)ribose: Ribose Deoxyribose ’ 3’ 5’ 3’

33 PCR Melting 96º, 30 sec Annealing ~55º, 30 sec Extension 72º, 30 sec 35 cycles Animation :

34 PCR Animation: PCR graph:

35 Gel electrophoresis DNA fragments are seperated using gel electrophoresis –Typically 1% argarose –Colored with EtBr or ZybrGreen (glows in UV light). –A DNA ”ladder” is used for identification of known DNA lengths. Gel picture: PCR setup: + -

36 The Sanger method of DNA sequencing Images: } Terminator X-ray sequenceing gel OH

37 Automated sequencing The major break-through of sequencing has happended through automation. Fluorescent dyes. Laser based scanning. Capillary electrophoresis Computer based base- calling and assembly. Images:

38 Handout exercise: ”base-calling” Handout: Chromotogram Groups of 2-3. Tasks: –Identify “difficult” regions –Identify “difficult” sequence stretches. –Try to estimate the best interval to use.

39 Biological data on computers The GenBank database File formats –FASTA –GenBank

40 NCBI GenBank GenBank is one of the main internaltional DNA databases. GenBank is hosted by NCBI: National Center for Biotechnology Information. GenBank has exists since The database is public - no restrictions on the use of the data within.

41 FASTA format >alpha-D ATGCTGACCGACTCTGACAAGAAGCTGGTCCTGCAGGTGTGGGAGAAGGTGATCCGCCAC CCAGACTGTGGAGCCGAGGCCCTGGAGAGGTGCGGGCTGAGCTTGGGGAAACCATGGGCA AGGGGGGCGACTGGGTGGGAGCCCTACAGGGCTGCTGGGGGTTGTTCGGCTGGGGGTCAG CACTGACCATCCCGCTCCCGCAGCTGTTCACCACCTACCCCCAGACCAAGACCTACTTCC CCCACTTCGACTTGCACCATGGCTCCGACCAGGTCCGCAACCACGGCAAGAAGGTGTTGG CCGCCTTGGGCAACGCTGTCAAGAGCCTGGGCAACCTCAGCCAAGCCCTGTCTGACCTCA GCGACCTGCATGCCTACAACCTGCGTGTCGACCCTGTCAACTTCAAGGCAGGCGGGGGAC GGGGGTCAGGGGCCGGGGAGTTGGGGGCCAGGGACCTGGTTGGGGATCCGGGGCCATGCC GGCGGTACTGAGCCCTGTTTTGCCTTGCAGCTGCTGGCGCAGTGCTTCCACGTGGTGCTG GCCACACACCTGGGCAACGACTACACCCCGGAGGCACATGCTGCCTTCGACAAGTTCCTG TCGGCTGTGTGCACCGTGCTGGCCGAGAAGTACAGATAA >alpha-A ATGGTGCTGTCTGCCAACGACAAGAGCAACGTGAAGGCCGTCTTCGGCAAAATCGGCGGC CAGGCCGGTGACTTGGGTGGTGAAGCCCTGGAGAGGTATGTGGTCATCCGTCATTACCCC ATCTCTTGTCTGTCTGTGACTCCATCCCATCTGCCCCCATACTCTCCCCATCCATAACTG TCCCTGTTCTATGTGGCCCTGGCTCTGTCTCATCTGTCCCCAACTGTCCCTGATTGCCTC TGTCCCCCAGGTTGTTCATCACCTACCCCCAGACCAAGACCTACTTCCCCCACTTCGACC TGTCACATGGCTCCGCTCAGATCAAGGGGCACGGCAAGAAGGTGGCGGAGGCACTGGTTG AGGCTGCCAACCACATCGATGACATCGCTGGTGCCCTCTCCAAGCTGAGCGACCTCCACG CCCAAAAGCTCCGTGTGGACCCCGTCAACTTCAAAGTGAGCATCTGGGAAGGGGTGACCA GTCTGGCTCCCCTCCTGCACACACCTCTGGCTACCCCCTCACCTCACCCCCTTGCTCACC ATCTCCTTTTGCCTTTCAGCTGCTGGGTCACTGCTTCCTGGTGGTCGTGGCCGTCCACTT CCCCTCTCTCCTGACCCCGGAGGTCCATGCTTCCCTGGACAAGTTCGTGTGTGCCGTGGG CACCGTCCTTACTGCCAAGTACCGTTAA (Handout)

42 GenBank format Originates from the GenBank database. Contains both a DNA sequence and annotation of feature (e.g. Location of genes). (handout)

43 GenBank format - HEADER LOCUS CMGLOAD 1185 bp DNA linear VRT 18-APR-2005 DEFINITION Cairina moschata (duck) gene for alpha-D globin. ACCESSION X01831 VERSION X GI:62724 KEYWORDS alpha-globin; globin. SOURCE Cairina moschata (Muscovy duck) ORGANISM Cairina moschata Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Archosauria; Aves; Neognathae; Anseriformes; Anatidae; Cairina. REFERENCE 1 (bases 1 to 1185) AUTHORS Erbil,C. and Niessing,J. TITLE The primary structure of the duck alpha D-globin gene: an unusual 5' splice junction sequence JOURNAL EMBO J. 2 (8), (1983) PUBMED COMMENT Data kindly reviewed (13-NOV-1985) by J. Niessing.

44 GenBank format - ORIGIN section ORIGIN 1 ctgcgtggcc tcagcccctc cacccctcca cgctgataag ataaggccag ggcgggagcg 61 cagggtgcta taagagctcg gccccgcggg tgtctccacc acagaaaccc gtcagttgcc 121 agcctgccac gccgctgccg ccatgctgac cgccgaggac aagaagctca tcgtgcaggt 181 gtgggagaag gtggctggcc accaggagga attcggaagt gaagctctgc agaggtgtgg 241 gctgggccca gggggcactc acagggtggg cagcagggag caggagccct gcagcgggtg 301 tgggctggga cccagagcgc cacggggtgc gggctgagat gggcaaagca gcagggcacc 361 aaaactgact ggcctcgctc cggcaggatg ttcctcgcct acccccagac caagacctac 421 ttcccccact tcgacctgca tcccggctct gaacaggtcc gtggccatgg caagaaagtg 481 gcggctgccc tgggcaatgc cgtgaagagc ctggacaacc tcagccaggc cctgtctgag 541 ctcagcaacc tgcatgccta caacctgcgt gttgaccctg tcaacttcaa ggcaagcggg 601 gactagggtc cttgggtctg ggggtctgag ggtgtggggt gcagggtctg ggggtccagg 661 ggtctgagtt tcctggggtc tggcagtcct gggggctgag ggccagggtc ctgtggtctt 721 gggtaccagg gtcctggggg ccagcagcca gacagcaggg gctgggattg catctgggat 781 gtgggccaga ggctgggatt gtgtttggaa tgggagctgg gcaggggcta gggccagggt 841 gggggactca gggcctcagg gggactcggg gggggactga gggagactca gggccatctg 901 tccggagcag gggtactaag ccctggtttg ccttgcagct gctggcacag tgcttccagg 961 tggtgctggc cgcacacctg ggcaaagact acagccccga gatgcatgct gcctttgaca 1021 agttcttgtc cgccgtggct gccgtgctgg ctgaaaagta cagatgagcc actgcctgca 1081 cccttgcacc ttcaataaag acaccattac cacagctctg tgtctgtgtg tgctgggact 1141 gggcatcggg ggtcccaggg agggctgggt tgcttccaca catcc //

45 FEATURES Location/Qualifiers source /organism="Cairina moschata" /mol_type="genomic DNA" /db_xref="taxon:8855" CAAT_signal TATA_signal precursor_RNA /note="primary transcript" exon /number=1 CDS join( , , ) /codon_start=1 /product="alpha D-globin" /protein_id="CAA " /db_xref="GI: " /db_xref="GOA:P02003" /db_xref="InterPro:IPR000971" /db_xref="InterPro:IPR002338" /db_xref="InterPro:IPR002340" /db_xref="InterPro:IPR009050" /db_xref="UniProt/Swiss-Prot:P02003" /translation="MLTAEDKKLIVQVWEKVAGHQEEFGSEALQRMFLAYPQTKTYFP HFDLHPGSEQVRGHGKKVAAALGNAVKSLDNLSQALSELSNLHAYNLRVDPVNFKLLA QCFQVVLAAHLGKDYSPEMHAAFDKFLSAVAAVLAEKYR" repeat_region /note="direct repeat 1" intron /number=1 repeat_region /note="direct repeat 1" exon /number=2 intron /number=2 exon /number=3 polyA_signal polyA_signal 1114 GenBank format - FEATURE section

46 Exercise: GenBank Work in groups of 2-3 people. The exercise guide is linked from the course programme. Read the guide carefully - it contains a lot of information about GenBank.


Download ppt "Introduktion til Bioinformatik Hold 01 Oktober 2010."

Similar presentations


Ads by Google