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
Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display 10-7
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