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Characterization of a full-length cDNA for rabbit factor X
U R Pendurthi1, K D Anderson, H L James
1Department of Biochemistry, University of Texas Health Center at Tyler 75710, USA.
Thrombosis Research
|March 15, 1997
Summary
Researchers isolated and characterized rabbit factor X (RFX) cDNA. This study reveals RFX shares high homology with human factor X, impacting activation through inhibitory peptides.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- Factor X is a crucial coagulation factor.
- Understanding species-specific Factor X provides insights into hemostasis and evolution.
Purpose of the Study:
- To isolate and characterize the complementary DNA (cDNA) for rabbit factor X (RFX).
- To analyze the structural and functional similarities between RFX and human factor X (HFX).
Main Methods:
- Polymerase Chain Reaction (PCR) amplification of a conserved region from rabbit genomic DNA.
- Northern blot analysis of rabbit liver RNA to detect RFX mRNA.
- cDNA library screening (lambda Zap II) from rabbit liver mRNA.
- DNA sequencing and amino acid sequence analysis.
Main Results:
- A 300 bp probe was generated for RFX detection.
- Northern blot confirmed a 1.6 kb mRNA species for RFX.
- The isolated RFX cDNA sequence includes regulatory regions, an open reading frame, and a poly(A) tail.
- The deduced amino acid sequence shows significant homology to HFX.
- Inhibitory peptides targeting HFX interaction sites partially inhibited RFX activation, with inhibition correlating to sequence homology.
Conclusions:
- The cDNA for rabbit factor X has been successfully isolated and characterized.
- Rabbit factor X shares substantial structural homology with human factor X.
- Functional studies using inhibitory peptides suggest conserved interaction sites between RFX and HFX.