武汉大学：《遗传学》课程教学课件（英文讲稿）Chapter 09B Genomes and proteomes-mod5-mod

Chapter9BGenomes and Proteomes10-1

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Sections to study1Large-scale genome mapping and analysis2 Major insights from the human and model organism genomesequences3Global analysis of genes and theirmRNAs4Globalanalysisofproteomes5 Repercussions of the human genome project and highthroughputtechnology10-2

Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display 10-2 Sections to study Sections to study 1 Large 1 Large-scale genome mapping and analysis scale genome mapping and analysis 2 Major insights from the human and model organism genome 2 Major insights from the human and model organism genome sequences sequences 3 Global analysis of genes and their mRNAs 3 Global analysis of genes and their mRNAs 4 Global analysis of proteomes 4 Global analysis of proteomes 5 Repercussions of the human genome project and high 5 Repercussions of the human genome project and high￾throughput technology throughput technology

The major advances that have transformed geneticsRecombinantHumanDNADNAGenomeMendel'stechnologystructureProjectdiscoveries186619531970s1990Genetics10-3

Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display 10-3 The major advances that have transformed genetics The major advances that have transformed genetics 1866 Mendel’s discoveries 1953 DNA structure 1990 Human Genome Project Genetics 1970s Recombinant DNA technology

1 Large-scale genome mapping and analysisGenome: The entire geneticChromosomeromatidChromatidchNuclnuinformation in a particularTolomerecell or organism.Genomics:The study of wholegenomes. It dedicated to theCelldevelopmentand applicationofmore effectivemappingsequencing,andcomputational toolsNA/doubi

Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display 10-4 1 Large-scale genome mapping and analysis  Genome: The entire genetic information in a particular cell or organism.  Genomics: The study of whole genomes. It dedicated to the development and application of more effective mapping, sequencing, and computational tools

Techniques forgenome mapping and sequencingCloningLibraryofDNAfragments500-1,000,000bpInsertintoone vectorHybridization Location of aparticular DNAsequencewithina library offragmentsPCRamplification DirectamplificationofaparticularregionofDNArangingfrom1oobpto>2okbDNA sequencingAutomated DNA sequencer using Sangermethod determines sequenceslooo bpat a time.Computational toolsPrograms for identifying matches between a particular sequence and a largepopulation of previously sequenced fragmentsPrograms foridentifying overlaps ofDNA fragmentsPrograms for estimating errorratesPrograms foridentifyinggenes in chromosomal sequences10-5

Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display 10-5 Techniques for genome mapping and sequencing Techniques for genome mapping and sequencing  Cloning Cloning  Library of DNA fragments 500 Library of DNA fragments 500 – 1,000,000 1,000,000 bp  Insert into one vector Insert into one vector  Hybridization Hybridization  Location of a particular DNA se Location of a particular DNA sequence within a library of fragme quence within a library of fragments  PCR amplification PCR amplification  Direct amplification of a particul Direct amplification of a particular region of DNA ranging from ar region of DNA ranging from 100 bp to > 20kb to > 20kb  DNA sequencing DNA sequencing  Automated DNA sequencer using Sang Automated DNA sequencer using Sanger method determines sequences er method determines sequences 1000 bp at a time. at a time.  Computational tools Computational tools  Programs for identifying matches Programs for identifying matches between a particular sequence a between a particular sequence and a large nd a large population of previously sequenced fragments population of previously sequenced fragments  Programs for identifying overlaps of DNA fragments Programs for identifying overlaps of DNA fragments  Programs for estimating error rates Programs for estimating error rates  Programs for identifying ge Programs for identifying genes in chromosomal sequences nes in chromosomal sequences

Vectors used for cloning large inserts for physical mappingYACs(yeastartificialchromosomes)Insertsize250-2,000kbBACs(bacterialartificialchromosomes)Insertsize100-300kbMore stableand easier to purifyfromhostDNA than YACs10-6

Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display 10-6 Vectors used for cloning large inserts for physical mapping Vectors used for cloning large inserts for physical mapping  YACs (yeast artificial chromosomes) (yeast artificial chromosomes)  Insert size 250 Insert size 250-2,000 kb 2,000 kb  BACs (bacterial artificial chromosomes) (bacterial artificial chromosomes)  Insert size 100 Insert size 100 – 300 kb  More stable and easier to purify from host DNA than More stable and easier to purify from host DNA than YACs

EcoRIcloning siteCEN4SUP4ARS1TRP1PYACURA3vectorAmporiTELTELTargetDNABamHIBamHlBamHl and EcoRl digestPartial EcoRl digestTELTRPARSCENTEL福URALigateURATELTELTRPARSCENVZ10-7

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Large-scale linkage maps serve as guides to whole genomesAFLP281AFLP201AFLP010AFLP003AFLP331 DNA Markerstrait ALinkedtoAAFLP057AFLP099AFLP291AFLP020AFLP458AFLP222DNAMarkerstraitBLLinkedtoBAFLP142AFLP033AFLP19910-8

Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display 10-8 Large-scale linkage maps linkage maps serve as guides to whole genomes serve as guides to whole genomes

Long-range physical mapsPhysical maps: a constellation of overlapping DNA fragments thatare ordered and oriented and span each of the chromosomes in agenome.STS (Sequence tagged sites):ShortDNA sequences that are alreadylocatedandamplifiedbyapairofPCRprimers.EachSTSrepresentsauniqueDNA sequenceinthegenome.STS1STS2BAC1BAC2STS3BAC3STS4BAC4STS5BAC5STS6BAC6STS7BAC7BAC810-9

Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display 10-9 Long-range physical maps physical maps Physical maps: a constellation of overlapping DNA fragments that are ordered and oriented and span each of the chromosomes in a genome.  STS (Sequence tagged sites): Short DNA sequences that are already (Sequence tagged sites): Short DNA sequences that are already located and amplified by a pair located and amplified by a pair of PCR primers. Each STS represe of PCR primers. Each STS represents a unique DNA sequence in the genome. unique DNA sequence in the genome

Locating cloned genes or markers on the chromosomeof a karyotype》BY11Complete set ofa3humanchromosomes川11%1格路216stained with9.2-7104Giemsa dye shows16RR14品品4bands.1316174Ddab)Ideograms（染色体308866.42G带图）showxChnidealized bandingpattern.888102innxxxxxxxnFig. 10.2 a, b22

Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display 10-10 Locating cloned genes or markers on the chromosome Locating cloned genes or markers on the chromosome of a karyotype karyotype a) Complete set of Complete set of human chromosomes chromosomes stained with stained with Giemsa dye shows bands. b) Ideograms ( Ideograms (染色体 G带图) show idealized banding idealized banding pattern. pattern. Fig. 10.2 a, b