A novel missense mutation responsible for factor VII deficiency in research Beagle colonies

J Thromb Haemost. 2006 Dec;4(12):2616-22. doi: 10.1111/j.1538-7836.2006.02203.x. Epub 2006 Sep 8.


Background: Canine factor VII (cFVII) deficiency, an autosomal recessive trait originally identified in research Beagles, is associated with a mild to moderate bleeding tendency.

Objective: Our aim was to identify and characterize the mutation causing cFVII deficiency.

Methods: In order to sequence the coding regions of the cFVII gene, we cloned the cFVII cDNA. Genomic DNA and plasma from FVII-deficient Beagles and obligate carriers were utilized.

Results: In all FVII-deficient dogs, we identified a single causative G to A missense mutation in exon 5, encoding the second epidermal growth factor-like domain, resulting in substitution of glycine 96 by glutamic acid, with plasma FVII coagulant activity of <or = 4% in affected Beagles. In vitro expression indicated that the majority (96%) of cFVII-G96E protein was retained intracellularly. In addition, analysis of purified recombinant wild-type and mutant cFVII proteins demonstrated reduced activity of the mutant (< 2%) compared with wild-type. Rotational thromboelastometry revealed a severe impairment of clotting activity in affected Beagles, and heterozygotes also exhibited changes in coagulation-based assays. Using a mutation-specific polymerase chain reaction/restriction digest that allows rapid identification of the G96E mutation, we surveyed a US research Beagle colony and identified a mutant allelic frequency of 31%.

Conclusions: We have identified a single causative mutation for cFVII deficiency that may have implications for pharmacotoxicologic research, because reduced FVII coagulant activity may alter hemostatic and/or cardiovascular endpoints in this commonly used animal species.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blood Coagulation
  • Cell Line
  • Cloning, Molecular
  • DNA Mutational Analysis
  • Dog Diseases / blood
  • Dog Diseases / genetics*
  • Dogs
  • Factor VII / genetics*
  • Factor VII / metabolism
  • Factor VII Deficiency / blood
  • Factor VII Deficiency / genetics*
  • Factor VII Deficiency / veterinary*
  • Gene Frequency
  • Heterozygote
  • Homozygote
  • Humans
  • Molecular Sequence Data
  • Mutation, Missense*
  • Polymerase Chain Reaction
  • Prothrombin Time
  • Recombinant Proteins / metabolism
  • Sequence Alignment
  • Sequence Analysis, Protein
  • Thrombelastography
  • Transfection


  • Recombinant Proteins
  • Factor VII