ANAT 322 Lecture Notes - Lecture 5: Ventromedial Nucleus Of The Hypothalamus, Viral Vector, Intron
3 Sep 2016
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Document Summary
Dna w/goi w/promoter (of gene or another) into fertilized egg pronucleus dna will randomly integrate into chromosome fertilized egg into pseudopregnant female some will be transgenic can create colony. Random integration could land in genomic desert, or transcribed site. Circadian clock molecular clock, 24h rhythms of gene expression. Can visualize rhythmicity by create transgenic animal w/promoter of period gene. Luciferase as reporter, convert luciferene w/emittance of photons record bioluminescence, direct readout of promoter activity. Diff tissues w/diff rhythms evidence for body rhythm in all tissues. Negative selection marker: cell will die when integrated into genome, in single crossover. Ko: goi knocked out in every cell type; not always wanted, if want to study function in specific tissue/cells. Remove exon necessary for gene function: put loxp upstream & downstream of goi by recombination low amount of cre recombinase to remove wrong loxps goi w/flanking.
