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Transgenic (Tg) 2-12H mice were engineered by incorporating the rearranged V(D)J segment derived from the 2-12H hybridoma. This variable immunoglobulin gene segment was cloned upstream of the Igha constant region (Cu) gene. The purified DNA construct was microinjected into blastocysts from (C57BL/6 × SJL)F1 crosses, which were subsequently implanted into pseudopregnant females. Two transgenic founder lines were established and backcrossed to either C57BL/6 or C.B-17 mice for a minimum of eight generations. The mixed-origin line has since been further backcrossed to C57BL/6 for thirteen generations. Transgene transmission was confirmed via polymerase chain reaction (PCR) analysis of tail DNA, utilizing a forward oligonucleotide primer (5'-CAGACTAAGGCCAAATATCAACTGAGAGA-3') matching a sequence 470 base pairs (bp) upstream of the 2-12 Vh exon and a reverse primer (5'-CAGGCTCCACCAGACCTCTCTAGA-3') complementary to a sequence downstream of Jh4.
Transgenic Vk8 mice were generated by microinjection of cloned functionally rearranged Vk8 L chain gene DNA into (C57BL/6 x SJL) F1 x F1 zygotes and transferred into pseudopregnant ICR/Halcr females. Inbred lines were established by backcrossing founder mice to BALB/c mice for six or more generations. The BALB/c background have been backcrossed to C57BL/6 thirteen generations. Mice containing the transgene were identified by polymerase chain reaction analysis of tail DNA using an oligonucleotide complementary to the Vk8 leader (5'-GGTACCTGTGGGACATTGTG- 3') and an oligonucleotide specific to the Vk8/Jk5 junction (5'- AGCACCGAACGTGAGAGG-3'). Mice containing the 2-12H transgene and mice containing the Vk8 transgene were bred to produce mice with both transgenes, referred to as 2-12h/Vk8 Tg mice.
Wild-type littermates
Colony Surveillance Program and Current Health 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 mmrrc@med.unc.edu 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.
The donor or their institution limits the distribution to non-profit institutions only.
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.
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