Strain Name
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C57BL/6N-Miftm1(MIF)Bcgen Ddttm1(DDT)Bcgen/Bcgen
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Common Name
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B-hMIF/hDDT mice
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Background
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C57BL/6N
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Catalog number
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121219
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Related Genes
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GIF, GLIF, MMIF; D-DT, DDCT, MIF-2, MIF2
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NCBI Gene ID
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4282; 1652
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Targeting strategy
Gene targeting strategy for B-hMIF/hDDT mice. The exons 1-3 of mouse Mif gene that encode the whole molecule is replaced by human counterparts in B-hMIF/hDDT mice. The genomic region of mouse Mif gene that encodes cytoplasmic portion is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric MIF expression is driven by endogenous mouse Mif promoter, while mouse Mif gene transcription and translation will be disrupted.
The exons 1-3 of mouse Ddt gene that encode the whole molecule is replaced by human counterparts in B-hMIF/hDDT mice. The genomic region of mouse Ddt gene that encodes cytoplasmic portion is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric DDT expression is driven by endogenous mouse Ddt promoter, while mouse Ddt gene transcription and translation will be disrupted.
Protein expression analysis
Strain specific MIF expression analysis in homozygous B-hMIF/hDDT mice by ELISA. Serum were collected from wild-type C57BL/6N mice (+/+) (n=2, famle, 7w) and homozygous B-hMIF/hDDT mice(H/H;H/H) (n=3, famle, 7w) stimulated with LPS in vivo (20 μg/mice, stimulation for 4 hours, i.p.), and analyzed by ELISA with species-specific MIF ELISA kit. Expression level of mouse and human MIF were analyzed by ELISA (mouse MIF ELISA: abcam, ab209885; human MIF ELISA: RD, DMF00B). Mouse MIF was detectable in wild-type mice. Human MIF was exclusively detectable in homozygous B-hMIF mice but not in wild-type mice.
Western blot analysis of DDT protein expression in homozygous B-hMIF/hDDT mice. Various tissue lysates were collected from homozygous B-hMIF/hDDT mice (H/H;H/H), and then analyzed by western blot with anti-DDT antibody(Abcam, ab115785). 40 μg total proteins were loaded for western blotting analysis. DDT was detected in liver and testis. (HO: homozygous B-hMIF/hDDT mice (H/H;H/H); +/+: wild-type C57BL/6N mice (+/+)).
mRNA expression analysis
Strain specific analysis of DDT mRNA expression in wild-type C57BL/6N mice and B-hMIF/hDDT mice by RT-PCR. Liver RNA were isolated from wild-type C57BL/6N mice (+/+) (n=1, famle, 7w) and homozygous B-hMIF/hDDT mice (H/H;H/H) (n=1, famle, 7w), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse and human DDT primers. Mouse Ddt mRNA was only detectable in wild-type mice but not in homozygous B-hMIF/hDDT mice. Human DDT mRNA was only detectable in homozygous B-hMIF/hDDT mice but not in wild-type mice.