cDNA cloning of rabbit thrombomodulin and characterization of gene expression in cardiovascular tissue.
Thrombomodulin (TM), a component of the protein C anticoagulant pathway, is critical for the maintenance of vascular thromboresistance. To facilitate the study of in vivo TM regulation, we cloned and sequenced the cDNA encoding full-length rabbit TM. Translation of the open reading frame predicts a 580 amino acid protein that contains a 19 amino acid signal peptide, one lectin-like and six EGF-like extracellular domains, a 23 amino acid transmembrane domain and a 36 amino acid cytoplasmic domain. In addition, there are three potential N-linked and six O-linked glycosylation sites. Comparison of the predicted rabbit TM protein with those of human, mouse and rat reveals 67-72% primary sequence conservation with identical domain homology. TM gene expression was quantified in rabbit cardiovascular tissue by real-time PCR using primers and probe based on the derived cDNA sequence and found to correlate with protein expression as determined by Western blot analysis.
Deming
Clayton B
CB
Division of Cardiology, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA.
Kim
Antony Y
AY
Bian
C E
CE
Regard
Jean B
JB
Rade
Jeffrey J
JJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004730:Endothelium, Vascular / Q000378:metabolism; D005786:Gene Expression Regulation; D007601:Jugular Veins / Q000637:transplantation; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism*; D011817:Rabbits; D051381:Rats; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D018180:Thrombomodulin / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors
D018076:DNA, Complementary; D018180:Thrombomodulin