Plasmid DNA pGL-PPRE5x-Ptk was linearized with NotI and SalI to generate a 3.1-kb fragment of PPRE transgene for the generation of transgenic mice following pronuclear microinjection of FVB oocytes. Of 18 offspring (F0), six tested positive for PPRE transgene by polymerase chain reaction (PCR) genotyping (primer-P 5′-AACTGCATAAGGCTATGAAGAGATACGCCC-3′ and primer-M 5′-TTAAAACCGGGAGGTAGATGAGATGTGACG-3′). All transgenic mice were crossed with wild-type F1 mice to yield PPRE heterozygous mice with the FVB genetic background.
