Cloning and sequencing of the canine and feline cardiac troponin I genes.
OBJECTIVE
To determine the gene sequences of canine and feline cardiac troponin I (cTnI), express the protein from the cloned gene in vitro, and validate the use of a commercial cTnI serum analyzer in these species via detection of the expressed protein or comparison of sequence homology.


METHODS
Samples of ventricular myocardium from 5 healthy adult mixed-breed dogs and 5 healthy adult domestic shorthair cats.


METHODS
The RNA was extracted from myocardial samples, and cDNA was synthesized via reverse transcriptase polymerase chain reaction and sequenced. The canine cDNA for the coding region was expressed in cell culture and analyzed by western blot and sandwich enzyme-linked immunosorbent assays.


RESULTS
Canine and feline cTnI genes were cloned and sequenced. Homology of the nucleotide and amino acid sequences of the canine and feline cTnI genes with human and rodent cTnI genes were high; the greatest homology was detected between canine and feline genes (95% and 96%, respectively). Recombinant canine cTnI protein was detected by a commercial serum cTnI analyzer and by western blot analysis.


CONCLUSIONS
Results indicated that commercial cTnI analyzers can be used to measure serum cTnI concentration from dogs and cats. Additionally, our preliminary characterization of the feline cTnI gene may facilitate further investigation of cTnI and its role in familial hypertrophic cardiomyopathy in cats.
Rishniw
Mark
M
Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
Barr
Stephen C
SC
Simpson
Kenny W
KW
Winand
Nena J
NJ
Wootton
Joyce A M
JA
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002415:Cats / Q000235:genetics*; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004285:Dogs / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D008969:Molecular Sequence Data; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013053:Spectrophotometry; D019210:Troponin I / Q000235:genetics*
D017931:DNA Primers; D018076:DNA, Complementary; D019210:Troponin I