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Cloning and sequence analysis of human calcyphosine complementary DNA
H El Housni1, A Radulescu, R Lecocq
1IRIBHN, Université Libre de Bruxelles, Faculté de Médecine, Brussels, Belgium.
Biochimica Et Biophysica Acta
|June 26, 1997
Summary
Calcyphosine, a calcium-binding protein, shows high sequence conservation between dogs and humans. However, human thyrocytes express significantly lower levels of calcyphosine mRNA and protein compared to canine thyrocytes.
Area of Science:
- Molecular Biology
- Biochemistry
- Comparative Genomics
Background:
- Calcyphosine, initially identified as thyroid protein p24, is a calcium-binding protein with four EF-hand domains.
- It was first characterized in dogs and is known as the R2D5 antigen in rabbits.
Purpose of the Study:
- To isolate and characterize the human counterpart of canine calcyphosine.
- To compare the sequence conservation and expression levels of calcyphosine between canine and human thyrocytes.
Main Methods:
- Isolation of human calcyphosine cDNA using a canine cDNA probe.
- Northern blot analysis to assess mRNA abundance.
- Western blot analysis to determine protein levels.
Main Results:
- Human calcyphosine cDNA was isolated and showed high nucleotide and amino acid sequence conservation with canine calcyphosine.
- The closest homologue identified was the crustacean CCBP-23 protein.
- Calcyphosine mRNA and protein levels were significantly lower in human thyrocytes compared to canine thyrocytes.
Conclusions:
- Calcyphosine is a highly conserved protein between canines and humans.
- Despite sequence conservation, there are significant species-specific differences in calcyphosine expression in thyroid cells.