Geneomics and Database Mining and Genetic Mapping DNA make RNA makes Protein and proteins do things…
Sequence databases change how we “do” biology
Mapping requires: The ability to measure traits Differences that segregate in populations Statistical and computational techniques to build maps in conventional and unconventional populations
What techniques can we use to measure genetic differences and similarities? RFLPs showing co-dominant inheritance in an F2
PCR-based CAPs, SNPs, SSRs……. Markers showing expected 7:1 segregation in an inbred backcross populations (BC2S5)
AFLPs showing dominant inheritance.
How to “mine” sequence data for: Genetic markers SSRs, SNPs, Conserved motifes How to analyze sequence data using distributed resources Identification of ORFs Identification of probable gene function more….
Saturated maps and targeted marker hunts Many crops and model species have saturated genetic maps Marker data can be accessed through specialized web sites (e.g. SGN for tomato http://www.sgn.cornell.edu) map position trait data (cross specific) Sequence of selected RFLPs
Detection of SNPs “in silico” Align BLAST two sequences ClustalX
Finding signal through the noise False positive The rate of PM < error True SNP
Sequence data opens up possibilities of designing markers in silico from conserved regions (e.g. resistance gene analogs).
New applications: Proteomics – from protein to gene
2-DE of low stringency fractions (soluble protein) Inoculated 2353 Un-inoculated 2353
2-DE of high stringency fractions (membrane bound)
From protein to gene O
From protein to gene From peptide fragments or amino acid sequence to DNA sequence is software driven. Example: Expert Protein Analysis System proteomics server of the Swiss Institute of Bioinformatics Protein Prospector UCSF Mass Spectrometry Facility
Before we use markers in the lab for a mapping exercise…… Introduction to public data-bases Basic DNA sequence manipulation (FASTA format, Sequence utilities, ORF finder, BLAST) Peptide Sequence searching Exercise (first hand experience)
http://www.oardc.ohio-state.edu/tomato francis.77@osu.edu