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Blood coagulation factor X mRNA encodes a single polypeptide chain containing a prepro leader sequence
Nucleic Acids Research
|June 11, 1984
Summary
Researchers cloned bovine factor X complementary DNA (cDNA) from a liver library. The cDNA sequence reveals factor X is synthesized as a precursor requiring multiple proteolytic steps for activation.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- Bovine factor X is a crucial protein in the coagulation cascade.
- Understanding its synthesis pathway is vital for hemostasis research.
Purpose of the Study:
- To clone and sequence the complementary DNA (cDNA) of bovine factor X.
- To elucidate the synthesis and processing of factor X.
Main Methods:
- Screening of a bovine liver cDNA library using synthetic oligonucleotides.
- Plasmid DNA isolation and restriction endonuclease digestion.
- DNA sequencing of the largest insert (pBX1).
Main Results:
- Five positive colonies containing bovine factor X cDNA inserts were identified.
- Plasmid pBX1 contained a 1530bp insert confirming successful cDNA cloning.
- Sequence analysis predicted a single-chain precursor with a prepro leader peptide.
Conclusions:
- Bovine factor X is synthesized as a prepro protein.
- At least five proteolytic cleavage events are required for mature factor Xa formation.