Basic Information
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Targeting strategy
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Gene targeting strategy for B-hHBEGF mice.
The exons 2~4 of mouse Hbegf gene that encode the extracellular domain were replaced by human HBEGF exons 2~4 in B-hHBEGF mice. -
Protein expression analysis
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Western blot analysis of HBEGF protein expression in homozygous B-hHBEGF mice. Various tissue lysates were collected from wild-type C57BL/6N (+/+) mice and homozygous B-hHBEGF mice (H/H), and then analyzed by western blot with anti-HBEGF antibody. 40μg total proteins were loaded for western blotting analysis. HBEGF were detected in kidney, liver and spleen. The antibodies showed cross-recognition.
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mRNA expression analysis
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Strain specific analysis of HBEGF mRNA expression in wild-type C57BL/6N mice and B-hHBEGF mice by RT-PCR. Heart and lung RNA were isolated from wild-type C57BL/6N mice (+/+) and homozygous B-hHBEGF mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse and human HBEGF primers. As the coding region is only about 630bp, the same primers were used to amplify both mouse Hbegf and human HBEGF gene. PCR products in heart and lung for human HBEGF were confirmed via Sanger Sequencing.
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Summary
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mRNA expression analysis:
Mouse Hbegf mRNA were detectable in heart and lung of wild-type C57BL/6N mice. Human HBEGF mRNA were detectable in B-hHBEGF mice, and were confirmed via Sanger Sequencing.Protein expression analysis:
Human and mouse HBEGF was both detectable on kidney, liver and spleen of WT mice and homozygous B-hHBEGF mice, as the antibody showed cross-recognition.
