Basic Information
-
Targeting strategy
-
Gene targeting strategy for B-hKIT/hKITLG mice.
The exons 2-7 of mouse Kitlg gene that encode the extracellular region were replaced by human KITLG exons 2-7 in B-hKIT/hKITLG mice.
A chimeric coding sequences of human KIT gene encoding the extracellular region and mouse Kit gene encoding the signal peptide, transmembrane region and the intracellular region were inserted in exon 2 of mouse Kit gene in B-hKIT/hKITLG mice.
-
mRNA expression analysis
-
Strain specific analysis of KITLG gene expression in wild-type mice and B-hKITLG mice by RT-PCR. Mouse KITLG mRNA was detectable in lung of wild-type mice (+/+). Human KITLG mRNA was detectable only in homozygous B-hKITLG mice (H/H), but not in wild-type mice.
-
Protein expression analysis
-
Strain specific KITLG expression analysis in homozygous B-hKITLG mice by ELISA. Serum was collected from wild-type mice (+/+) and homozygous B-hKITLG mice (H/H), and analyzed by ELISA with species-specific KITLG ELISA kit. Mouse KITLG was detectable in wild-type mice. Human KITLG was exclusively detectable in homozygous B-hKITLG mice but not in wild-type mice.
Strain specific KIT expression analysis in homozygous B-hKIT mice by flow cytometry. Bone marrow were collected from wild-type mice (+/+) and homozygous B-hKIT mice (H/H),and analyzed by flow cytometry with species-specific anti-KIT antibody. Mouse KIT was detectable in wild-type mice. Human KIT was exclusively detectable in homozygous B-hKIT mice but not in wild-type mice.
-
Summary
-
mRNA expression analysis:
Mouse KITLG mRNA was detectable in lung of wild-type mice (+/+). Human KITLG mRNA was detectable only in homozygous B-hKITLG mice (H/H), but not in wild-type mice.
Protein expression analysis:
Human KITLG was exclusively detectable in homozygous B-hKITLG mice but not in wild-type mice. Human KIT was exclusively detectable in homozygous B-hKIT mice but not in wild-type mice.
