We tested this using a mouse Rpa70 fragment containing the OBN, DBD-A and DBD-B domains (residues 1 to 431; thereafter Rpa70NAB).
In brief, 4.7 kb of mouse genomic fragment containing the second exon of the Cxcr4 gene was floxed by two loxP sites containing neomycin-resistant (Neor) gene, and the diphtheria toxin A (DT-A) gene was inserted to the upstream of the 5' arm for negative selection.
For the generation of CTMP mutant mice, a mouse genomic DNA fragment containing exons 2 and 3 was cloned into the pBluescript vector and a Not1 site was generated in exon 2. A ∼5-kb IRES-lacZ-Neo cassette was inserted into the NotI site, which introduced a translational frame shift.
In this mouse strain, the fragment containing the calmodulin-binding domain of NOS2 was replaced by the neomycin resistance gene [9].
A mouse Bcor cDNA fragment containing sequences from within exon 4 to within exon 7 was used to screen the RPCI-22 129S6/Sv EvTac Bac library (Stratagene) [63], and positive BAC clones were used to clone Bcor genomic sequence.
The BAC clone, designated BAC1, carried a 114 kb mouse genomic DNA fragment containing two class I odorant receptor genes [ 10].
The GPT element inserted into all mice was a fragment containing 1.1 kb upstream of the golli transcription start site plus 0.2 kb downstream into the first exon of the MBP gene.
This site was blunted, and a blunted DNA fragment containing the mouse GFAP promoter, which was prepared from pGF1L (Miura et al. 1990) with HindIII, was inserted.
The Sca1-MafB vector was generated as follows: The 1.3-kb EcoRI- EcoRI fragment, containing the mouse MafB cDNA, was inserted into the ClaI site of the pLy6 vector (Miles et al, 1997) to generate the Sca1-MafB vector.
To generate pMSCV-ΔNp63 α, a PCR fragment containing the mouse ΔNp63α gene using murine pcDNA3-ΔNp63 α as a template was subcloned into the retroviral vector pMSCV-puro (Clontech, Palo Alto, CA, USA) digested with BglII and HpaI.
