研人类基因组学.ppt

Prerequiste of human genome sequencing: genetic and physical maps Genetic Mapping – based on recombination frequency (expressed as cM); Key word : co-segregation Physical Mapping – actual molecular distance in nucleotide base pairs (expressed as bp, kb, or mb) Key word: contig Genetic maps are important crutches for physical maps 2011-10-25 向光盛 长江大学医学院 Genetic Markers for Mapping Polymorphic markers for genetic mapping. RFLPs – restriction fragment length polymorphisms SSRs – simple sequence repeats (also called microsatellites) For High-resolution physical mapping STSs – sequence tagged sites ESTs – expressed sequenced tags 2011-10-25 向光盛 长江大学医学院 Fragment of a human genetic map One map unit = one centimorgan (cM) = 1% recombination between loci 2011-10-25 向光盛 长江大学医学院 Physical maps 2011-10-25 向光盛 长江大学医学院 Ways to create a genetic maps:Analysis of large human pedigrees The CEPH Family Panel (Centre du Etude Polymorphisme Humain) – 40 nuclear families 10 are French families: 27 are Utah Mormon pedigrees: 2 are Venezuelan Huntingtons pedigrees: 1 is an Old Order Amish pedigree: (has bipolar affective disorder segregating). The total number of individuals in the panel is 520, with an average sibship size of 8. 2011-10-25 向光盛 长江大学医学院 Typical task of CEPH based research To integrate new polymorphic marker into human genetic map: To PCR given marker in DNA from members of CEPH families To compare segregation patterns of given marker with segregation patterns of other known markers (already mapped) To conclude genetic location of marker of interest as co-segregated with known marker 2011-10-25 向光盛 长江大学医学院 1. Somatic cell hybrids 2. Radiation hybrids 3. Enrichments of starting DNA for library construction - chromosome flow-sorting; - microdissection; 4. Contig construction from genomic fragments - BAC, YAC, PAC….. - cosmids…. Ways to create a physical maps: 2011-10-25 向光盛 长江大学医学院 Somatic cell hybrids cgil.uoguelph.ca/ QTL/