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Related Experiment Videos

Cloning and expression of human tissue factor cDNA.

K L Fisher1, C M Gorman, G A Vehar

  • 1Department of Cardiovascular Research, Genentech, Inc., South San Francisco, CA 94080.

Thrombosis Research
|October 1, 1987
PubMed
Summary

Researchers cloned and expressed human tissue factor complementary DNA (cDNA). This membrane protein is crucial for initiating blood coagulation and fibrin clot formation.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Hematology

Background:

  • Tissue factor is a key membrane protein initiating blood coagulation.
  • It interacts with factor VII to activate factors IX and X, driving fibrin clot formation.

Purpose of the Study:

  • To clone and express the complementary DNA (cDNA) for human tissue factor.
  • To characterize the encoded protein and its function.

Main Methods:

  • Complementary DNA (cDNA) cloning and sequencing of human tissue factor.
  • Northern blot hybridization to detect tissue factor mRNA.
  • Transfection of mammalian cells with an expression vector containing the cDNA.
  • Assay of expressed tissue factor activity using purified factors VII and X.

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Main Results:

  • The human tissue factor cDNA encodes a 263-amino acid protein with a signal peptide.
  • The protein has a hydrophobic membrane-anchoring domain and no significant homology to other proteins.
  • Tissue factor mRNA was detected in multiple tissues, including adipose and adrenal glands.
  • Transfected cells produced active tissue factor.

Conclusions:

  • The successful cloning and expression of human tissue factor cDNA provide a tool for further research.
  • The findings confirm the role of tissue factor in blood coagulation and its tissue distribution.