Generalized lacZ expression with the ROSA26 Cre reporter strain

P Soriano - Nature genetics, 1999 - nature.com
Nature genetics, 1999nature.com
Fig. 1 ROSA 26 targeting. a, Top, restriction map of the locus. PCR primers from ROSA26
flanking (5− CCTAAAGAAGAGGCTGTGCTTTGG− 3) and splice acceptor (5−
CATCAAGGAAACCCTGGACTACTG− 3) sequences were used to amplify an approximately
1.2-kb diagnostic fragment (grey arrowheads). The probe used for Southern-blot analysis is
shown as a shaded box. LoxP sites are indicated by black arrowheads. Only EcoRV sites
are indicated for pROSA26-R. b, Left, Southern-blot analysis of targeted clone (1− 8) and …
Fig. 1 ROSA 26 targeting. a, Top, restriction map of the locus. PCR primers from ROSA26 flanking (5− CCTAAAGAAGAGGCTGTGCTTTGG− 3) and splice acceptor (5− CATCAAGGAAACCCTGGACTACTG− 3) sequences were used to amplify an approximately 1.2-kb diagnostic fragment (grey arrowheads). The probe used for Southern-blot analysis is shown as a shaded box. LoxP sites are indicated by black arrowheads. Only EcoRV sites are indicated for pROSA26-R. b, Left, Southern-blot analysis of targeted clone (1− 8) and wild-type (WT) DNA digested with EcoRV; 1C and 2C are populations of ES clones 1 and 2 transiently transfected with PGKCrebpA (a gift of M. Komada) showing the expected shorter targeted fragment due to deletion of the neo segment. Right, only the shorter EcoRV fragment is seen in offspring also expressing Cre (lanes 6, 7) in contrast with the reporter allele alone (lane 2). ROSA26-R embryos could be genotyped by PCR (approximately 500-bp wild-type and (approximately 250-bp mutant fragments) using three oligonucleotides: 5− AAAGTCGCTCTGAGTTGTTAT− 3, 5− GCGAAGAGTTTGTCCTCAACC− 3 and 5− GGAGCGGGAGAAATG GATATG− 3.
nature.com