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6 Cards in this Set

  • Front
  • Back
Transgenic mouse
-multiple species
-insert foriegn DNA randomly into pronucleus of fertilized egg, then implant egg into foster mom
-exam w/tail biopsies-PCR or south blot
-usually-ectopic misexpression(in cells where not normally expressed)
-Or overexpression
-random insertion=variable expression
Transgenic mouse Preperation
1)Plasmid containing-promotor(high levels or tissue specific),transgene,termination/polyA makes stable mRNA
2)fertilized egg
3)foster mom
4)other mice to mate positive pups
Transgenic mouse applications
A)Analysis of gene function through overexpression
B)Animal models of human disease(extension of (A),models diseases assoc. with overexpression)
C)Production of foriegn proteins
Knock-out mice
-mice only
-targeted insertion in ES cells
-disrupt specific genes to reveal func.
-exam-by linking hair color to gene
Targeted insertion production
1)Plasmid and targeting sequence must be homologous recombination-pos/neg selection
2)corectly targeted ES cells microinjected into blastocysts removed from donor female
3)put into foster pseudopregnant mom
4)mate chimeric mice to wild-type mice
Target insertion applications
A)Knock-out suble mutations,even point mutations to interfere with aspects of gene func.
B)Gene knock-in=replacement or alteration of endogenous gene
C)manipulate gene copy number to raise or lower level of production
D)intro of reporter gene for studies of promotor function