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Adopted from PMID:10402668: The Adh7 gene targeting vector was linearized with NotI and introduced by electroporation into R1 mouse embryonic stem cells using established methodology. Cells were subjected to both, positive selection with G418 and negative selection with ganciclovir, to enrich for cells incorporating the construct by homologous recombination. Genomic DNA was isolated from surviving cell clones and screened by Southern blot analysis using HindIII digestion to identify cells in which a portion of the Adh7 gene was deleted. The external DNA probe for Southern blot screening consisted of a 1.5-kb XbaI fragment containing exon 9 of Adh7. In Adh7+/- clones, the wild-type HindIII fragment was 4.0 kb, and the mutant HindIII fragment was 5.8 kb. The truncated mutant Adh7 gene lacks the promoter and exons 1–6 containing the coding region for amino acid residues 1–275. Karyotype analysis was performed on Southern blot-positive clones to identify clones having a normal karyotype for further use. Adh7+/- mutant embryonic stem cells were microinjected into C57BL/6 blastocysts, which were then implanted into pseudopregnant females. This resulted in chimeric mice having a representation of cells derived from the 129/Sv embryonic stems cells as identified by patches of hair with the dominant agouti coat color of the 129/Sv strain. Several chimeric male mice were mated to wild-type female Black Swiss mice and germline transmission of the introduced embryonic stem cells was identified by offspring with a complete agouti coat color. Southern blot analysis of tail DNA from agouti offspring was used to identify individuals heterozygous for the Adh7 mutation, i.e., carrying both the 4.0- and the 5.8-kb HindIII fragments described above. Heterozygous Adh7+/- mutant mice were mated to produce homozygous Adh7-/- mutant mice also identified by Southern blot analysis of tail DNA. Adh7 mutant and wild-type mice were maintained on Purina basal diet 5755, unless otherwise stated. Northern blot analysis with an Adh7 cDNA probe was performed on mouse tissues using 10 µg of total RNA as previously described. Western blotting with polyclonal antibodies against mouse Adh1, Adh3, and Adh7 was accomplished using 20 µg of total protein from mouse tissues as reported.
Deltour L, Foglio MH, Duester G. Impaired retinol utilization in Adh4 alcohol dehydrogenase mutant mice. Dev Genet. 1999;25(1):1-10. (Medline PMID: 10402668)
Cryo-recovered strains distributed by the MMRRC at JAX are shipped to the customer from the Pathogen & Opportunistic-Free Animal Room G200 - see https://www.jax.org/jax-mice-and-services/customer-support/customer-service/animal-health/health-status-reports.
Limited quantities of breeder mice (recovered litter) are available from a cryoarchive; recovered litter usually available to ship in 3 to 4 months.
Cryopreserved material may be available upon request, please inquire to csmmrrc@jax.org for more information.
Distribution of this strain requires submission of the MMRRC Conditions of Use (COU). A link to the COU web form will be provided via email after an order has been placed; the form should be completed then or the email forwarded to your institutional official for completion.
Additional charges may apply for any special requests. Shipping costs are in addition to the basic distribution/resuscitation fees. Information on shipping costs and any additional charges will be provided by the supplying MMRRC facility.
1 The distribution fee covers the expense of rederiving mice from a live mouse; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.
2 An aliquot contains a sufficient number of embryos (in one or more vials or straws and based on the transfer success rate of the MMRRC facility) to transfer into one to three recipients. The MMRRC makes no guarantee concerning embryo transfer success experienced in the recipient investigator's laboratory. Neither gender nor genotype ratios are guaranteed.
3 An aliquot is one straw or vial with sufficient sperm to recover at least one litter of mice, as per provided protocols, when performed at the MMRRC facility. The MMRRC makes no guarantee concerning the success of these procedures when performed outside the MMRRC facilities.
4 The distribution fee covers the expense of resuscitating mice from the cryo-archive; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.