Identification and characterization of new variants of three associated SNPs and a microsatellite in the TSH recpetor gene which are usefull for genetic studies.

SEBASTIAN A ESPERANTE, CARINA M RIVOLTA, MARIELA CAPUTO, ROGELIO GONZALEZ SARMIENTO, HECTOR MANUEL TARGOVNIK

Molecular and Cellular Probes

Academic Press

Lugar: England; Año: 2008 vol. 22 p. 281 - 286

0890-8508

The purpose of the present work was to characterize g.IVS5-69C>T, g.IVS6+13C>T and c.561C>T SNPs and the (CT)n microsatellite in the TSHR gene for genetic analysis. Exons 6 and 7 of the TSHR gene, including the falnkin intronic sequences, were screened for the presence of g.IVS5-69C>T, g.IVS6+13C>T and c.561C>T by SSCP. We found genetic association between the three SNPs and a total of three different Haplotypes were observed. Two were homozygous blocks, g.IVS5-69T,/g.IVS6+13G/c.561C (Haplotype TGC, 3,3%) and g.IVS5-69C,/g.IVS6+13A/c.561T (Haplotype CAT, 75%). Every individual who was heterozygous for g.IVS5-69C>T, was equally heterozygous for g.IVS6+13C>T and c.561C>T (Haplotype CAT/TGC, 21,7%).The (CT)n microsatellite, localized in intron 7 of the TSHR gene was amplified by PCR and the labeled products were separeted in a polyacrylamide denaturing sequencing gel/ Three variable numbers of CT motif were identified, two previously reporter (CT)6 and (CT)8 and one unreporter (CT)9. The construction and expression of the hybrid minigenes using pSPL3 and alfa globin fibronectin EDB (pTB) showed that the (CT)n microsatellite does not interfere with exon 8 definition and processing in vitro.In conclusion g.IVS5-69C>T, g.IVS6+13C>T and c.561C>T and (CT)n microsatellite are informative polymorphic markers and can be used in linkage studies in families with germ line THSR mutations or autoinmunity thyroid diseases.