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The targeting vector for generating the inducible A1R
knock-out allele was constructed using plasmid
pB-Not-XhoMaxi (pMaxi), which contains the major coding exon
of the A1R [homologous to human exon 6; a gift from B.
Johansson. A loxP site along with HindIII and BamHI sites
were inserted into the EcoR1 site 5 to the exon. The 4.5 kb
fragment from plasmid pGB128 containing the cytosine
deaminase and neomycin resistance genes was inserted into
the NcoI site 3 to exon 6. This fragment was flanked by loxP
sites to allow removal of the cassette. All three loxP sites
in the final targeting vector were in the same 5 to 3
orientation. A 1.6 kb fragment from plasmid
pGKneobpAlox2PGKDTA containing the diphtheria toxin gene was
inserted into the XhoI site of pMaxi. The final targeting
vector was linearized with NotI and transfected into J1
embryonic stem (ES) cells derived from 129SvJ mice.
Genomics - Transgenes
National Institutes of Health (NIH)
Curation Flag Information
New comment to be added:
Hyperinsulinemic clamp order #30460 Nov 2020
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Financial support for this work was provided by the NIDDK Mouse Metabolic Phenotyping Centers (MMPC-
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