Description
- TGF-alpha is a member of the EGF family of cytokines that are synthesized as transmembrane precursors with soluble forms released by proteolytic cleavage. Membrane-bound proTGF-alpha is biologically active and plays a role in mediation of cell-cell adhesion and in juxtacrine stimulation of adjacent cells.
- TGF-alpha binds and activates the EGF receptor. It induces fibroblast proliferation, epithelial development, keratinocyte migration, and angiogenesis.
- Expression of TGF-alpha is widespread in tumors and transformed cells. TGF-alpha is also expressed in normal tissues during embryogenesis and in adult tissues, including pituitary, brain, keratinocytes and macrophages.
- TGFA is most highly expressed in heart, lung, kidney, small intestine, thymus and lymph nodes.
- The exons 3~4 of mouse Tgfa gene was replaced by human TGFA exons 3~4 in B-hTGFA mice.
Targeting strategy
Gene targeting strategy for B-hTGFA mice. The exons 3~4 of mouse Tgfa gene was replaced by human TGFA exons 3~4 in B-hTGFA mice.
Experimental design
Strain specific TGFA expression analysis in wild-type C57BL/6 mice and homozygous humanized B-hTGFA mice by ELISA. Serum was collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTGFA mice (H/H) (n=3, 10-week-old). Protein expression level of TGFA was analyzed by ELISA. Human TGFA was exclusively detectable in homozygous B-hTGFA mice
Strain specific TGFA expression analysis in wild-type C57BL/6 mice and homozygous humanized B-hTGFA mice by ELISA. BMDM was collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTGFA mice (H/H), stimulated with 10 μg/mL ConA for 5d. Protein expression level of TGFA from cell culture supernatant was analyzed by ELISA. Human TGFA was exclusively detectable in homozygous B-hTGFA mice.
mRNA expression analysis
Strain specific analysis of TGFA mRNA expression in wild-type C57BL/6 mice and B-hTGFA mice by RT-PCR. Lung RNA were isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hTGFA mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human TGFA primers. Mouse Tgfa mRNA was detectable only in wild-type C57BL/6 mice. Human TGFA mRNA was detectable only in homozygous B-hTGFA mice but not in wild-type mice.
