Insertion of a pathogenic mutation into a yeast artificial chromosome containing the human amyloid precursor protein gene.
The genetic modelling of human disease would be considerably facilitated if pathogenic mutations could be inserted into transgenes which were then expressed in an appropriate manner. Yeast artificial chromosomes (YACs), when used as transgenes appear to direct expression with the correct temporal and spatial distribution. Here we demonstrate that it is comparatively straightforward to introduce pathogenic mutations into such YACs by the use of the 'pop-in, pop-out' procedure, by inserting an Alzheimer-causing mutation (APP717Val-->lle) into an APP-containing YAC which has previously been used as a transgene. The significance of these procedures for the modelling of human disease is discussed.
Duff
K
K
Suncoast Alzheimer's Disease Laboratories, Department of Psychiatry, University of South Florida, Tampa 33613, USA.
McGuigan
A
A
Huxley
C
C
Schulz
F
F
Hardy
J
J
D000544:Alzheimer Disease / Q000235:genetics; D000682:Amyloid / Q000235:genetics*; D001483:Base Sequence; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D005821:Genetic Techniques; D006801:Humans; D008957:Models, Genetic; D008969:Molecular Sequence Data; D017354:Point Mutation; D000072002:Prion Proteins; D011328:Prions; D011498:Protein Precursors / Q000235:genetics*; D011995:Recombination, Genetic
D000682:Amyloid; D017931:DNA Primers; C512656:PRNP protein, human; D000072002:Prion Proteins; D011328:Prions; D011498:Protein Precursors