Copyright, ©, 2002, John Wiley & Sons, Inc.,Karp/CELL & MOLECULAR BIOLOGY 3E Footprints and Shadows Looking for Functional Pieces Within Genomes
Copyright, ©, 2002, John Wiley & Sons, Inc.,Karp/CELL & MOLECULAR BIOLOGY 3E
Copyright, ©, 2002, John Wiley & Sons, Inc.,Karp/CELL & MOLECULAR BIOLOGY 3E Footprints vs. Shadows Footprints –sequences conserved in “distant” organisms –works less well than you might think –many alignments not functional (about 40%) –typical comparison: mouse and man limited to mammalian conserved sequence primate conserved sequence would be missed
Copyright, ©, 2002, John Wiley & Sons, Inc.,Karp/CELL & MOLECULAR BIOLOGY 3E Footprints vs. Shadows Shadows –sequences conserved in “similar” organisms –not very effective when comparing two organisms –high fraction of pairwise similarity –multiple simultaneous comparisons better primates MORE evolutionary distance than mouse/man
Copyright, ©, 2002, John Wiley & Sons, Inc.,Karp/CELL & MOLECULAR BIOLOGY 3E
Copyright, ©, 2002, John Wiley & Sons, Inc.,Karp/CELL & MOLECULAR BIOLOGY 3E CETP LXR apoB plas 4 exons and flanking regions
Copyright, ©, 2002, John Wiley & Sons, Inc.,Karp/CELL & MOLECULAR BIOLOGY 3E Apo(a) promoter E = exon C = conserved, N = not 9 = TATA, 10 = HNF-alpha
Copyright, ©, 2002, John Wiley & Sons, Inc.,Karp/CELL & MOLECULAR BIOLOGY 3E Gel retention (shift) assay
Copyright, ©, 2002, John Wiley & Sons, Inc.,Karp/CELL & MOLECULAR BIOLOGY 3E Deletion/Transfection Assay of apo(a)