Molecular cloning, characterization and expression of cDNA for rabbit brain tissue factor

Area of Science:

• Biochemistry
• Molecular Biology
• Cardiovascular Research

Background:

• Tissue factor (TF) initiates blood coagulation and is found in atherosclerotic plaques.
• TF may trigger thrombosis following plaque rupture, a risk in conditions like acute myocardial infarction and angioplasty.
• An established atherosclerotic rabbit model is available for studying angioplasty outcomes.

Purpose of the Study:

• To isolate and characterize rabbit cDNA encoding mature tissue factor (TF).
• To provide a foundational step towards developing inhibitors of TF.
• To investigate the functional activity of expressed rabbit TF.

Main Methods:

• Isolation and characterization of rabbit TF cDNA.
• Sequence comparison of rabbit, human, and mouse TF cDNA.
• Expression of rabbit TF cDNA in E. coli.
• Assessment of procoagulant activity and antibody blocking.

Main Results:

• Rabbit TF cDNA was successfully isolated and characterized.
• Rabbit TF cDNA shows significant sequence similarity to human and mouse TF.
• Expressed rabbit TF exhibits comparable procoagulant activity to native TF.
• Polyclonal antibodies against rabbit TF effectively blocked TF activity.

Conclusions:

• The characterized rabbit TF cDNA provides a tool for further research.
• The findings support the development of TF inhibitors for thrombotic conditions.
• This work is crucial for understanding TF's role in atherosclerosis and angioplasty complications.