Basic Information
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Description
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The SLC4A1 (Band 3) protein is the most abundant integral protein of human erythrocyte membranes and functions as the major anion transporter polypeptide of erythrocytes. SLC4A1 is a 90-100 kDa protein, its microheterogeneity is ascribed mainly to the structural heterogeneity of the carbohydrate moiety. The C-terminal portion of the molecule spans the membrane several times, and is responsible for anion transport. The N-terminal portion, comprising almost half of the polypeptide chain of SLC4A1, is located inside the red cell and interacts with cytoskeletal and cytoplasmic proteins such as ankyrin.
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Targeting strategy
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Gene targeting strategy for B-hSLC4A1 mice. The exons 2-20 of mouse Slc4a1 gene that encode signal peptide, extracellular domain, transmembrane domain, cytoplasmic region and 3’UTR are replaced by human counterparts in B-hSLC4A1 mice. The promoter and 5’UTR region of the mouse gene are retained. The human SLC4A1 expression is driven by endogenous mouse Slc4a1 promoter, while mouse Slc4a1 gene transcription and translation will be disrupted.
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mRNA expression analysis
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Strain specific analysis of SLC4A1 mRNA expression in wild-type C57BL/6N mice and B-hSLC4A1 mice by RT-PCR. Kidney RNA were isolated from wild-type C57BL/6N mice (+/+) and homozygous B-hSLC4A1 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human SLC4A1 primers. Mouse Slc4a1 mRNA was detectable in wild-type mice. Human SLC4A1 mRNA was detectable only in homozygous B-hSLC4A1 mice(H/H) but not in wild-type mice.
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Protein expression analysis
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Strain specific hCD233/SLC4A1 expression analysis in homozygous B-hSLC4A1 mice(H/H) by flow cytometry. Blood were collected from wild-type C57BL/6N mice(+/+) and B-hSLC4A1 mice(H/H) , and analyzed by flow cytometry with anti-human CD233/SLC4A1 antibody (Miltenyi biotec, 130-117-825). Human CD233/SLC4A1 was detectable in homozygous B-hSLC4A1 mice.
